bluuebunny/crossref_metadata_2025 huggingface dataset:
The impacts of Japanese direct investment in China on the Sino‐Japanese bilateral trade
Authors: Yu Jinping , Zhao Wenjun
Published: October 2008

Purpose

The economic linkages between China and Japan have been strengthened through both trade and Japanese direct investment in China for past decades. The purpose of this paper is to investigate the impacts of Japanese direct investment in China on the Sino‐Japanese bilateral trade.

Design/methodology/approach

An index, RRCA, was used to illustrate the changes of relative comparative advantage of major products, and AR(p) models used to examine the effects of Japanese FDI on both Chinese exports and Chinese imports.

Findings

This paper shows that Japanese direct investment in China has contributed not only to the increase of Chinese exports to Japan, but also to the increase of Chinese imports from Japan. This suggests that that the relations between Japanese direct investment in China and the bilateral trade are complementary.

Research limitations/implications

The sample size used in the empirical studies in this paper is very small. In addition, the studies focus only on the effects of FDI on trade while the effects of trade on FDI are neglected.

Originality/value

This paper provides evidence that Japanese direct investment in China is an important determinant factor of the rapid growth of the bilateral trade. It also shows that the appreciation of RMB against Japanese Yen is associated with a decrease in Chinese trade surplus in the bilateral trade.
Dynamic changes in comparative advantage: Japan “flying geese” model and its implications for China
Authors: Widodo Tri
Published: October 2008

Purpose

The purpose of this paper is to examine the Japan flying geese (FG) model and its implications for China.

Design/methodology/approach

Data on exports and imports three‐digit the standard international trade clasification (SITC) Revision 2 from UN‐COMTRADE are employed. An analytical tool namely “products mapping” is made by combining two fundamental variables derived from the FG model. Revealed symmetric comparative advantage (RSCA) index and trade balance index are applied.

Findings

The paper provides evidence of the existence of FG pattern. Unskilled labor‐intensive industries and human capital‐intensive industries have clearly shown the FG pattern in East Asia. China has very high comparative advantage in those industries.

Research limitations/implications

The classification of industries is a crucial issue. This paper applies the broader classification of industries based on factor intensity rather than end use. Further researches on more specific industries might give detailed explanation.

Originality/value

The paper examines the position of East Asian countries in the FG model.
Chinese distribution practitioners' attitudes towards Italian quality foods
Authors: Huliyeti Hasimu , Marchesini Sergio , Canavari Maurizio
Published: October 2008

Purpose

Food products' ultimate destination is intended for the general public, but those who supply the international markets are usually composed of a relatively small group of businessmen including international buyers, purchasing agents and importers. Understanding the motivation behind their purchasing decisions and the perceptions they have of the source countries can be considered a key factor in developing a successful marketing approach. This paper focuses on understanding the position of such a powerful minority towards imported Italian, high‐quality agro‐food products in China, as well as the current communication, marketing, strategic and organizational approaches of Italian producers, especially compared to other European competitors.

Design/methodology/approach

An exploratory approach based on qualitative interviews is adopted, with the aim of highlighting Chinese perceptions regarding Italian food suppliers' relevant strengths and weaknesses. The primary data were collected through personal interviews with 25 key informants in Shanghai, Beijing and Guangzhou.

Findings

Survey results indicated that while Italian products are appreciated, current market shares remain inferior to other countries due to inadequate marketing strategies and low investments in promotion. Price sensitivity of Chinese consumers, less‐established distribution channels and competition with local products hindered further the market penetration.

Originality/value

At the best of our knowledge there is no prior research concerning the gatekeepers attitudes towards quality food in the Chinese market. The paper provides a useful insight into a topic that is relevant togathers of marketing intelligence and planners of marketing strategy for China's market access in the rapidly changing and highly competitive food trade environment.
Function simulation of FDI, foreign trade and regional GDP in China
Authors: Ying Liu , Riming Cui
Published: October 2008

Purpose

The purpose of this paper is to simulate the function of foreign trade, foreign direct investment (FDI) and regional gross domestic product (GDP) in China, explore how these two variables affect regional GDP together and provide evidence to export‐led growth (ELG) and FDI‐led growth.

Design/methodology/approach

Artificial neural network (ANN) is introduced in the model. This nonlinear and adaptive computation obtains a three‐dimension function that is different from linear models.

Findings

New evidence was found for ELG and FDI‐led growth with data of 28 regions in China in the period of 1994‐2005. The simulation reveals that with foreign trade and FDI scale varying, marginal GDP in different Chinese regions is positive. Because of the nonlinear system, a wave pattern of marginal GDP was found and an optimal scale of foreign trade and FDI for Chinese regions. Results in the simulation also indicate the possibility of economic deconcentration in some Chinese regions.

Originality/value

New evidence is provided for ELG and FDI‐led growth. Different from conventional methods, ANN model as a nonlinear system is introduced in the study in which optimal scale of foreign trade and FDI for Chinese regions is obtained.
Special government/business relationship: the case of Taiwanese Businessmen Association in mainland China
Authors: Lee Chun‐Yi
Published: October 2008

Purpose

This paper focuses on the relationship between spontaneous business groups, such as the Taiwanese Businessmen Association (TBA), and the Chinese government.

Design/methodology/approach

In analysing the relationship between the TBAs and the Chinese government, this paper focuses on three cities, Tianjin, Kunshan and Dongguan. Furthermore it observes Taiwanese investment in mainland China from 1987 to 2004, in three sub‐periods: 1987‐1993, 1994‐1999, 2000‐2004. Statism (e.g. rational choice) appears to be a valid theoretical framework.

Findings

It was found that the Chinese government's changing interaction with Taiwanese businesses always suited their best interests at the time, which proves that the Chinese government applied rational module in interacting with Taiwanese businesses.

Research limitations/implications

Rational choice is not a comprehensive approach in political science, but it is the most suitable approach to analyse the interaction between the Chinese government and Taiwanese businesses.

Practical implications

The first TBA was established in Beijing in 1990; nowadays there are over 100 TBAs in mainland China. It has been argued that TBAs can be seen as the emergence of a sort of civil society in mainland China; to others, they merely reflect the strong will of the Chinese government to attract foreign investment.

Originality/value

This paper demonstrates that Taiwanese businesses is a security asset for the Chinese government in the cross‐Strait relationship.
Operationalization of corporate entrepreneurship and its performance implications in China
Authors: Wang Yonggui , Zhang Xiao
Published: October 2008

Purpose

It is intended in this paper to develop a reliable and valid measurement of corporate entrepreneurship (CE) and identify the key dimensions based on evidence of the largest transitional and fast growing economy, China, given that no agreement has been reached and much less is know about the generalizability of related research findings in emerging economies.

Design/methodology/approach

The eight‐step procedure for scale development was followed in this study, which is based on questionnaire survey and statistical analysis.

Findings

A four‐dimension measurement model for CE was developed and validated. Furthermore, the authors also find that not all dimensions of CE have statistically significant impacts on firm performance in China.

Originality/value

Nowadays, more and more studies have been conducted on entrepreneurship, entrepreneurial orientation, intrapreneurship or CE. However, no agreement has been reached about the key dimensions of CE and much less is know about the generalizability of related research findings in emerging economies. This paper is intended to bridge these gaps.
Channel performance in China: a study of distribution centers in Fujian Province
Authors: O'Neill Peter , José Scavarda Annibal , Yang Zhenhua
Published: October 2008

Purpose

This research seeks to undertake a comparative study of Chinese distribution centers (DCs) with UK DCs in order to explore the relationship between the supply chain, logistics and distribution functions, and the gap between the countries. The study has been undertaken in light of Chinese corporate standardization programs in distribution channels. The main goal of the standardization programs has been to create efficient supply chains to be able to control the flow of information, capital, product and other resources. The focus of the study was Fujian Province where the standardization program has meant that functional areas such as marketing and distribution are measured by key performance indicators (KPI), under specific evaluation keys. The study highlights benchmarked KPIs where supply chain entrepreneurship can be brought to bear.

Design/methodology/approach

The method used in this research is quantitative via a small scale sample. Questionnaires were sent to Chinese DCs in Fujian Province with additional quantitative information sought through telephone follow‐up.

Findings

The data have been analyzed and compared to UK DCs for inventory level, order lead‐times, customer segmentation, value‐added activities, and floor area utilization. DCs are a long way from internationally competitive best practice across complex product categories. This provides opportunity for entrepreneurial third party inventory storage and management. DCs also exhibits low transportation efficiency due to low vehicle loading factors, also affecting DC order lead‐time efficiency and value‐added activities. There is opportunity here for entrepreneurial activity in offering efficient (lean) and responsive (agile) third party transport services. There is also evidence of poor process management implying opportunity for the application of entrepreneurial consulting capability to resource and reengineer end‐end process chains.

Research limitations/implications

The research generalisability is limited due its small scale sample in a single Chinese province.

Practical implications

Despite a modernization program the implications of this study are that Chinese DCs are not ready for traditional international competition, but in the short‐term entrepreneurs may be able to design sustainable lean and agile supply chains that are at least as good as those in the UK.

Originality/value

The study's originality is its comparative benchmark of Chinese with UK DCs.
Acquisition of resources, formal organization and entrepreneurial orientation of new ventures
Authors: Fang Niu , Yuli Zhang , Hongzhi Xue
Published: October 2008

Purpose

The purpose of this paper is to assess the impact of the acquisition of resources and formal organization on entrepreneurial orientation (EO) of new ventures.

Design/methodology/approach

The research was conducted using an original data set of 199 new ventures in China. The data were analyzed through regressing models.

Findings

It was found that new ventures experiencing difficulty in acquiring resources have higher levels of EO; formal organization is associated with the proactiveness dimension of EO significantly and positively.

Research limitations/implications

First, the measures of EO were mainly used to study established firms. The measures of EO may need to be modified to take into account new ventures' uniqueness. Second, this study focused on new ventures operating in a specific geographical region, China.

Practical implications

New ventures should build their formal management systems so that they can integrate as organic organizations to engage in entrepreneurial activities. New ventures lack regulations and their development mainly depends on entrepreneurs. The formal regulations can help new ventures better use and integrate the power of the whole organization systems to identify and explore external opportunities.

Originality/value

Research on EO has focused on established firms while the EO of new ventures is seldom studied. These findings indicate the uniqueness of new ventures and contribute to a better understanding of their behaviors.
Entrepreneurship and strategy in China: why “Porter's five forces” may not be
Authors: Wang Wei , Chang Peter P.
Published: October 2008

Purpose

Challenging the suitability of “Porter's five forces” as a guide to strategy and entrepreneurship in China and in the global age at large, this paper aims to present a new “five forces” model based on Sun Tzu's timeless strategic wisdom, with entrepreneurship and “collaborative innovation” right at its heart.

Design/methodology/approach

The arguments and the new model are developed, drawing on results from a survey among entrepreneurs and executives operating in China, the direct experience of top entrepreneurs, authoritative information from publishers like the Financial Times and Business Week, the consensus at the 2008 Annual Conference of the World Economic Forum, and Sun Tzu's timeless strategic insights.

Findings

The survey among entrepreneurs and executives operating in China shows that “Porter's five forces” has had little impact on business practice in China. One author, a serial entrepreneur, has considered ren shi qian or three Ps (people, project and penny) as the strategic forces driving his ventures in China. Going beyond the narrow, extremely competitive focus of Porter's model, the new “five forces” model, consisting of business purpose, business location, business climate, business organisation and business leader, is shown to determine business success. And, as the central strategic force, business leaders have to embrace entrepreneurship and “collaborative innovation” with their hearts to navigate any challenging economic waters.

Originality/value

Freeing business practitioners and teachers/students from the dogma of “Porter's five forces” established 30 years ago, this paper will help them identify what really determines business success in the new age.
The Cashmere Collection: launching a new brand
Authors: Lightfoot William S. , Cohen Corine
Published: October 2008

Purpose

The purpose of this paper is to provoke further research and analysis into what it takes to successfully start and grow a new global venture, which sources a product in China, and resells it in Europe and the USA.

Design/methodology/approach

This paper explores the early stages of the establishment of a new venture, and a new brand. The venture – Zelective, was founded by two MBA students working on an applied research project that was then turned into a company located in the Principality of Monaco. Importing their cashmere clothing from Inner Mongolia, China, this small firm has enjoyed significant success by designing a line of clothing that is colorful, sexy, and distinctive, while also executing a sales and marketing strategy that has increased their brands awareness, and consequently, their sales to some top tier retailers in Europe.

Findings

The next challenge for the firm is to expand their efforts, while maintaining the provocative edge.

Originality/value

This case provides insight into the challenges of starting up a new venture which sources product from China. Students have a series of questions which are designed at having them research the opportunities for Zelective moving forward, helping them determine the firms growth strategy. This case can be used in a wide range of context, from marketing, sales, and brand management, to strategy and logistics.
The major shift towards user‐centred innovation
Authors: von Hippel Eric , Jin Chen
Published: October 2008

Purpose

The purpose of this paper is to highlight the nature and policy implications of the shift from producer‐centred product and service design to innovation by the user, specifically internet‐supported collaborative design by user communities.

Design/methodology/approach

An overview of the importance of innovation by individual lead users is followed by a description of collaborative innovation by user communities. The authors then focus on policies, which need to be re‐examined in the light of the shift from producer innovation to collaborative user innovation. The authors conclude by making some suggestions specifically pertinent to Chinese government policy, in a region, which is emerging as a major player in global innovation.

Findings

An important form of user innovation–Internet‐supported collaborative design of new products and services by user communities–is likely to progressively supplant producer product and service design in many fields. This shift will require changes in governments' innovation policies, for example, with respect to intellectual property rights (IPRs).

Originality/value

In line with emerging research findings, this paper takes a user‐centred view of the innovation process. It proposes applicable strategies, which will enhance the effectiveness of this novel innovation paradigm in a Chinese context.
Reflections on China and a six‐dimensional framework for managing research and development
Authors: Sofo Francesco
Published: October 2008

Purpose

There are continuing efforts by Chinese institutions to improve their approach to the management of research and development (R&D) within the national and global arenas. The purpose of this paper is to propose a framework for managing R&D in China.

Design/methodology/approach

The framework is derived from a review of two OECD reports, three books and 15 recently published articles in scholarly journals (1977‐2008) all of which are mainly related to the structure and organisation of research in foreign‐owned and Chinese research institutes.

Findings

In this paper, issues in the literature are identified and arranged in themes in order to discover key lessons about the management of R&D. The result of the analysis is a hybrid six‐dimensional model incorporating open innovation, connecting with the world, knowledge management, a public approach, balanced development and communities of inquiry.

Research limitations/implications

The framework emphasises a base of shrewd economic expansion supported by intelligent fiscal policies used as a basic building block that can be emulated within R&D in China and more broadly.

Practical implications

Practical applications include a structure for cross‐cultural analysis of R&D management to identify the mix of key elements and how this might augment success, build global empathy, foster deeper cooperation in R&D management and create inclusive competitive advantage.

Originality/value

This paper provides a new framework emanating from the synthesis of elements identified in the literature as being important to R&D management that can be applied by Chinese and foreign owned firms at a global level.
The effect of information technology and knowledge management capability on R&D process performance
Authors: Ma Huimin , Peng Yibing , Shi Yongjiang
Published: October 2008

Purpose

The main purpose of this paper is to develop the theoretical links and empirically examine the association between IT and R&D performance, and their study is to answer management's questions: how should IT resources be organized and managed to enhance the process performance?

Design/methodology/approach

The approach is consistent with the process perspective to the question of IT business value, but this approach is extend. A basic premise of this paper is that the knowledge management (KM) capability could be a critical mediator between IT capability and process performance.

Findings

The results indicate that an organization's KM capability is dependent on IT capabilities, and variation in business process performance is explained by KM capability.

Research limitations/implications

On the one hand, control variables were not included in this study. In fact, firm size might influence process performance. On another hand, the empirical analysis was conducted in the context of one specific process in the manufacturing industry; thus, the generalizability of the results may be limited.

Practical implications

The results indicated that IT managers have to clearly understand the strategic thrust of the organization and institute mechanisms to ensure that IT capabilities are channeled toward the areas important to the organization.

Originality/value

This study contributes to the IT literature by introducing KM capability as a critical mediator between IT capabilities and business process performance, this study improves our understanding of the true business value of IT.
Understanding the dynamism in Hong Kong's innovation environment
Authors: Sharif Naubahar , Baark Erik
Published: October 2008

Purpose

The authors' intention is to demonstrate how adopting a more precise interpretation of the term “institution” as used in the innovation studies literature provides a better understanding of the dynamic nature of an innovation system. The paper therefore focuses less on organisations and other key actors in Hong Kong's innovation system and more on the laws, rules, procedures, business practices, etc. that facilitate and govern such activity.

Design/methodology/approach

The paper adopts the term “innovation environment” to denote the institutional setting within which innovative activity is carried out in Hong Kong and reviews key elements of the innovation environment to note trends towards innovation‐oriented changes. Data and other descriptive information that informs the treatment of the innovation environment are based on interviews of diverse actors associated with Hong Kong's innovation system as well as research into an interdisciplinary body of literature.

Findings

The authors find that although there has been little evidence of substantial progress in the development of Hong Kong's innovation system, key elements of the innovation environment exhibit trends in the direction of a more robust use of innovation‐related resources to drive economic growth in the territory.

Originality/value

Apart from its focus on Hong Kong's innovation system, the paper advances innovation studies by demonstrating that its methodology of examining the innovation environment – the institutional setting in which key organisational actors carry out innovative activity – reveals the dynamism of an innovation system that might not emerge from a focus on organisations and other key actors.
Strategic quality management in an internationalised university department: a case study
Authors: Ratnatunga Janek
Published: October 2008

Purpose

In the early 1990s, a number of controversial and radical initiatives were introduced by the Australian Government that had a compounded impact on higher education in general, and for the teaching of accounting studies in particular. The impacts of these initiatives have now lasted well over a decade. The purpose of this paper is to address this situation.

Design/methodology/approach

This paper uses a case‐study approach to study the quality management objectives and strategies of an accounting department, to survive and prosper in this changing academic environment, initially as an independent entity in the period 1990‐1996, and thereafter from 1997 to 2006 as a merged entity with another accounting department in the same university that was an internal competitor in the earlier period.

Findings

It is demonstrated that the department was largely successful in climbing from a position of significant under‐achievement amongst its peers in 1990, to one of strength in terms of both research performance and entrepreneurship by 1994. However, continual re‐structuring of the department by the university has resulted in a loss of synergy and a decline in the latter periods of the study.

Practical implications

This case is of particular interest to educators who are coping with the issues of “balance” between teaching and research, and to those interested in seeing how a department implemented a comprehensive quality management programme largely in keeping with the framework provided by the Higher Education Council.

Originality/value

The value of the paper is that it provides many useful insights on many diverse issues to those universities and their departments which wish to operate in a globalised environment.
Assessment of students' perception towards developing a course in environmental accounting
Authors: Das Niladri , Sen Mitali , Pattanayak J.K.
Published: October 2008

Purpose

The purpose of this study is to assess the students' perception about the importance of incorporating environmental reporting practices, standards, and other issues in Management Accounting course.

Design/methodology/approach

Environmental issues provide a unique, timely, and important focus for developing a course in Indian management pedagogy, which shall help in demonstrating the usefulness of environmental accounting information in supporting corporate environmental strategies while assessing environmental performance. Assessment measurements that may capture the impact of reformed academic programs are necessary to judge the success and failure of management education reforms. This paper carries out such an assessment process by offering a course on environmental accounting as an intervention, to a sample of 28 management students enrolled in a higher educational institution in India imparting management education. A questionnaire is administered to these students before and after the intervention in order to understand the difference of their perception level.

Findings

The results suggests a significant difference in the perception level of students about the utility of incorporating environmental reporting practices in Management Accounting course before and after the administration of the reformed program.

Originality/value

The paper is unique in its attempt to understand the Indian management students' perception of knowledge and usefulness with environmental accounting in their regular course, from which a more detailed evaluation can be derived. Further work based on this preliminary finding may be used to develop a proposed environmental accounting course framework for management students in India.
Forward performance measurement and management integrated frameworks
Authors: Taticchi Paolo , Balachandran Kashi R.
Published: October 2008

Purpose

In recent years, performance measurement and management (PMM) has received much attention from researchers and practitioners. Despite the growing use of PMM systems, companies experience difficulty in implementing such systems, with consequent risk of partial benefits or total goal failure. The literature on PMM is quite vast, but only few of the models address the problem in its entirety, while many other works focus on specific issues related to PMM. The purpose of this paper is to analyze the state of the art of PMM models and propose an integrated framework as a base for performance measurement and management design.

Design/methodology/approach

The evolution of the literature on PMM models and frameworks is highlighted starting from the development of the last twenty years. Further, the characteristics raised in the literature are merged so as to identify the milestones of an integrated performance measurement and management system. Based on it, an integrated framework is proposed as a base for a cohesive PMM design.

Findings

The framework integrates five systems: a performance system, a cost system, a capability evaluation system, a benchmarking system and a planning system.

Research limitations/implications

Though the proposed framework is a starting point for performance measurement and management design, it provides important guidelines for successful implementations of PMM initiatives inside companies.

Practical implications

The paper elaborates on the findings in the literature through a review and explores how the framework proposed might be implemented and improved.

Originality/value

The framework is based on the belief that PMM study requires an intensive and deep comprehension of the business in focus, which begins with a complete analysis of all the key activities in the company and their related drivers. Accordingly, the framework proposed starts with a defining “which” information should be analyzed, “how” they should be processed and “how” they should be integrated for generating valuable information to facilitate managers' decision‐making processes.
An improved theory of constraints
Authors: Ray Amitava , Sarkar Bijan , Sanyal Subir Kumar
Published: October 2008

Purpose

The primary aim in this paper is to develop and demonstrate a theory of constraints (TOC) model in which constraint resource prevents the throughput of the organization.

Design/methodology/approach

In this paper, the authors propose an integrated model by combining Laplace criterion and TOC into a single evaluation model in a multiproduct constraint resource environment. A case study is illustrated to demonstrate the effectiveness of this model. The outsourcing decision model compares three alternatives: standard cost accounting, standard theory‐of‐constraints, and our own solution.

Findings

The numerical results show that the new approach is superior to Standard cost accounting and Theory of Constraints and presents a more realistic state of optimum allocation of resources and measures the performance of the model.

Research limitations/implications

This research is limited to the production processes that do not have multiple constraints.

Originality/value

This is the first time that the integrated model comprising of Laplace‐TOC model has been used to maximize the product throughput. Instead of calculating $return per constraint minute, this method decides the priority of product that maximizes the product throughput in the constraint resource environment. It makes a significant contribution to the manufacturing Organization where one can compare the financial performance of the Organization by selecting the right decision model.
Symmetrical dialogue in investor relations
Authors: Hassink Harold , Bollen Laury , de Vries Meinderd
Published: October 2008

Purpose

Previous research shows that symmetrical dialogue between companies and their investors through e‐mail does not match the quality of the asymmetrically provided information on investor relations websites. This paper seeks to further examine the quality of symmetrical dialogue and especially whether sender identity affects the corporate e‐mail handling performance.

Design/methodology/approach

A “mystery sender” approach is used to test whether companies answer similar e‐mails from two different stakeholder groups: investors and journalists. The corporate replies were evaluated for responsiveness, timeliness and relevance.

Findings

Overall responsiveness from the companies in the sample is low, and e‐mails from journalists turn out to get significantly fewer and less relevant replies than e‐mails from investors. Thus, the idea of sender identity affecting the way companies respond to e‐mail inquiries is empirically supported.

Practical implications

Stakeholders of listed companies should not rely too much on symmetrical dialogue by e‐mail given the high probability of non‐response, especially for journalists. Companies offering the possibility of symmetrical communication need to critically evaluate their information system to achieve better results in handling e‐mail from stakeholders and to make sure that certain stakeholder groups are not discriminated.

Originality/value

This is one of the first papers that present evidence on how corporate investor relation departments respond to e‐mails from different types of stakeholder groups. The paper contributes to a better understanding of the variables that drive the quality of symmetrical dialogue between companies and stakeholders groups.
Novel Method for Selection of Antimicrobial Peptides from a Phage Display Library by Use of Bacterial Magnetic Particles
Authors: Tanaka Tsuyoshi , Kokuryu Yoriko , Matsunaga Tadashi
Published: October 2008

Antimicrobial peptides were isolated from a phage display peptide library using bacterial magnetic particles (BacMPs) as a solid support. The BacMPs obtained from “
Magnetospirillum magneticum
” strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. BacMPs are easily purified from a culture of magnetotactic bacteria by magnetic separation. Approximately 4 × 10
10
PFU of the library phage (complexity, 2.7 × 10
9
) was reacted with BacMPs. The elution of bound phages from BacMPs was performed by disrupting its membrane with phospholipase D treatment. Six candidate peptides, which were highly cationic and could bind onto the BacMP membrane, were obtained. They exhibited antimicrobial activity against
Bacillus subtilis
but not against
Escherichia coli
and
Saccharomyces cerevisiae
. The amino acid substitution of the selected peptide, KPQQHNRPLRHK (peptide 6-7), to enhance the hydrophobicity resulted in obvious antimicrobial activity against all test microorganisms. The present study shows for the first time that a magnetic selection of antimicrobial peptides from the phage display peptide library was successfully achieved by targeting the actual bacterial inner membrane. This BacMP-based method could be a promising approach for a high-throughput screening of antimicrobial peptides targeting a wide range of species.
Purification and Gene Cloning of α-Methylserine Aldolase from
Ralstonia
sp. Strain AJ110405 and Application of the Enzyme in the Synthesis of α-Methyl-
l
-Serine
Authors: Nozaki Hiroyuki , Kuroda Shinji , Watanabe Kunihiko , Yokozeki Kenzo
Published: October 2008

By screening microorganisms that are capable of assimilating α-methyl-
dl
-serine, we detected α-methylserine aldolase in
Ralstonia
sp. strain AJ110405,
Variovorax paradoxus
AJ110406, and
Bosea
sp. strain AJ110407. A homogeneous form of this enzyme was purified from
Ralstonia
sp. strain AJ110405, and the gene encoding the enzyme was cloned and expressed in
Escherichia coli
. The enzyme appeared to be a homodimer consisting of identical subunits, and its molecular mass was found to be 47 kDa. It contained 0.7 to 0.8 mol of pyridoxal 5′-phosphate per mol of subunit and could catalyze the interconversion of α-methyl-
l
-serine to
l
-alanine and formaldehyde in the absence of tetrahydrofolate. Formaldehyde was generated from α-methyl-
l
-serine but not from α-methyl-
d
-serine,
l
-serine, or
d
-serine. α-Methyl-
l
-serine synthesis activity was detected when
l
-alanine was used as the substrate. In contrast, no activity was detected when
d
-alanine was used as the substrate. In the α-methyl-
l
-serine synthesis reaction, the enzymatic activity was inhibited by an excess amount of formaldehyde, which was one of the substrates. We used cells of
E. coli
as a whole-cell catalyst to express the gene encoding α-methylserine aldolase and effectively obtained a high yield of optically pure α-methyl-
l
-serine using
l
-alanine and formaldehyde.
Generation of Large Chromosomal Deletions in Koji Molds
Aspergillus oryzae
and
Aspergillus sojae
via a Loop-Out Recombination
Authors: Takahashi Tadashi , Jin Feng Jie , Sunagawa Misao , Machida Masayuki , Koyama Yasuji
Published: October 2008

We established a technique for efficiently generating large chromosomal deletions in the koji molds
Aspergillus oryzae
and
A. sojae
by using a
ku70
-deficient strain and a bidirectional marker. The approach allowed deletion of 200-kb and 100-kb sections of
A. oryzae
and
A. sojae
, respectively. The deleted regions contained putative aflatoxin biosynthetic gene clusters. The large genomic deletions generated by a loop-out deletion method (resolution-type recombination) enabled us to construct multiple deletions in the koji molds by marker recycling. No additional sequence remained in the resultant deletion strains, a feature of considerable value for breeding of food-grade microorganisms. Frequencies of chromosomal deletions tended to decrease in proportion to the length of the deletion range. Deletion efficiency was also affected by the location of the deleted region. Further, comparative genome hybridization analysis showed that no unintended deletion or chromosomal rearrangement occurred in the deletion strain. Strains with large deletions that were previously extremely laborious to construct in the wild-type
ku70
+
strain due to the low frequency of homologous recombination were efficiently obtained from Δ
ku70
strains in this study. The technique described here may be broadly applicable for the genomic engineering and molecular breeding of filamentous fungi.
Aerobic Fermentation of
d
-Glucose by an Evolved Cytochrome Oxidase-Deficient
Escherichia coli
Strain
Authors: Portnoy Vasiliy A. , Herrgård Markus J. , Palsson Bernhard Ø.
Published: October 2008

Fermentation of glucose to
d
-lactic acid under aerobic growth conditions by an evolved
Escherichia coli
mutant deficient in three terminal oxidases is reported in this work. Cytochrome oxidases (
cydAB
,
cyoABCD
, and
cbdAB
) were removed from the
E. coli
K12 MG1655 genome, resulting in the ECOM3 (
E. coli c
ytochrome
o
xidase
m
utant) strain. Removal of cytochrome oxidases reduced the oxygen uptake rate of the knockout strain by nearly 85%. Moreover, the knockout strain was initially incapable of growing on M9 minimal medium. After the ECOM3 strain was subjected to adaptive evolution on glucose M9 medium for 60 days, a growth rate equivalent to that of anaerobic wild-type
E. coli
was achieved. Our findings demonstrate that three independently adaptively evolved ECOM3 populations acquired different phenotypes: one produced lactate as a sole fermentation product, while the other two strains exhibited a mixed-acid fermentation under oxic growth conditions with lactate remaining as the major product. The homofermenting strain showed a
d
-lactate yield of 0.8 g/g from glucose. Gene expression and in silico model-based analyses were employed to identify perturbed pathways and explain phenotypic behavior. Significant upregulation of
ygiN
and
sodAB
explains the remaining oxygen uptake that was observed in evolved ECOM3 strains.
E. coli
strains produced in this study showed the ability to produce lactate as a fermentation product from glucose and to undergo mixed-acid fermentation during aerobic growth.
Modulation of Thiol-Disulfide Oxidoreductases for Increased Production of Disulfide-Bond-Containing Proteins in
Bacillus subtilis
Authors: Kouwen Thijs R. H. M. , Dubois Jean-Yves F. , Freudl Roland , Quax Wim J. , van Dijl Jan Maarten
Published: October 2008

Disulfide bonds are important for the correct folding, structural integrity, and activity of many biotechnologically relevant proteins. For synthesis and subsequent secretion of these proteins in bacteria, such as the well-known “cell factory”
Bacillus subtilis
, it is often the correct formation of disulfide bonds that is the greatest bottleneck. Degradation of inefficiently or incorrectly oxidized proteins and the requirement for costly and time-consuming reduction and oxidation steps in the downstream processing of the proteins still are major limitations for full exploitation of
B. subtilis
for biopharmaceutical production. Therefore, the present study was aimed at developing a novel in vivo strategy for improved production of secreted disulfide-bond-containing proteins. Three approaches were tested: depletion of the major cytoplasmic reductase TrxA; introduction of the heterologous oxidase DsbA from
Staphylococcus carnosus
; and addition of redox-active compounds to the growth medium. As shown using the disulfide-bond-containing molecule
Escherichia coli
PhoA as a model protein, combined use of these three approaches resulted in secretion of amounts of active PhoA that were ∼3.5-fold larger than the amounts secreted by the parental strain
B. subtilis
168. Our findings indicate that
Bacillus
strains with improved oxidizing properties can be engineered for biotechnological production of heterologous high-value proteins containing disulfide bonds.
DNA Microarray Analysis of the Heat Shock Transcriptome of the Obligate Intracytoplasmic Pathogen
Rickettsia prowazekii
Authors: Audia Jonathon P. , Patton Mary C. , Winkler Herbert H.
Published: October 2008

Here we present the first oligonucleotide DNA microarray analysis of global gene expression changes in the obligate intracytoplasmic pathogen
Rickettsia prowazekii
using temperature upshift as a model stress condition, and we describe a methodology for isolating highly purified rickettsial RNA. In toto, 23 transcripts were significantly increased by temperature upshift (≥2.0-fold;
P
< 0.05), and no transcripts demonstrated reproducible decreases. Array results for three heat shock-inducible mRNAs were confirmed using quantitative reverse transcription-PCR.
Relationship between Sublethal Injury and Microbial Inactivation by the Combination of High Hydrostatic Pressure and Citral or
tert
-Butyl Hydroquinone
Authors: Somolinos Maria , García Diego , Pagán Rafael , Mackey Bernard
Published: October 2008

The aim was to investigate (i) the occurrence of sublethal injury in
Listeria monocytogenes
,
Escherichia coli
, and
Saccharomyces cerevisiae
after high hydrostatic pressure (HHP) treatment as a function of the treatment medium pH and composition and (ii) the relationship between the occurrence of sublethal injury and the inactivating effect of a combination of HHP and two antimicrobial compounds,
tert
-butyl hydroquinone (TBHQ) and citral. The three microorganisms showed a high proportion of sublethally injured cells (up to 99.99% of the surviving population) after HHP. In
E. coli
and
L. monocytogenes
, the extent of inactivation and sublethal injury depended on the pH and the composition of the treatment medium, whereas in
S. cerevisiae
, inactivation and sublethal injury were independent of medium pH or composition under the conditions tested. TBHQ alone was not lethal to
E. coli
or
L. monocytogenes
but acted synergistically with HHP and 24-h refrigeration, resulting in a viability decrease of >5 log
10
cycles of both organisms. The antimicrobial effect of citral depended on the microorganism and the treatment medium pH. Acting alone for 24 h under refrigeration, 1,000 ppm of citral caused a reduction of 5 log
10
cycles of
E. coli
at pH 7.0 and almost 3 log
10
cycles of
L. monocytogenes
at pH 4.0. The combination of citral and HHP also showed a synergistic effect. Our results have confirmed that the detection of sublethal injury after HHP may contribute to the identification of those treatment conditions under which HHP may act synergistically with other preserving processes.
Staphylococcus aureus
-Fibronectin Interactions with and without Fibronectin-Binding Proteins and Their Role in Adhesion and Desorption
Authors: Xu Chun-Ping , Boks Niels P. , de Vries Joop , Kaper Hans J. , Norde Willem , Busscher Henk J. , van der Mei Henny C.
Published: October 2008

Adhesion and residence-time-dependent desorption of two
Staphylococcus aureus
strains with and without fibronectin (Fn) binding proteins (FnBPs) on Fn-coated glass were compared under flow conditions. To obtain a better understanding of the role of Fn-FnBP binding, the adsorption enthalpies of Fn with staphylococcal cell surfaces were determined using isothermal titration calorimetry (ITC). Interaction forces between staphylococci and Fn coatings were measured using atomic force microscopy (AFM). The strain with FnBPs adhered faster and initially stronger to an Fn coating than the strain without FnBPs, and its Fn adsorption enthalpies were higher. The initial desorption was high for both strains but decreased substantially within 2 s. These time scales of staphylococcal bond ageing were confirmed by AFM adhesion force measurement. After exposure of either Fn coating or staphylococcal cell surfaces to bovine serum albumin (BSA), the adhesion of both strains to Fn coatings was reduced, suggesting that BSA suppresses not only nonspecific but also specific Fn-FnBP interactions. Adhesion forces and adsorption enthalpies were only slightly affected by BSA adsorption. This implies that under the mild contact conditions of convective diffusion in a flow chamber, adsorbed BSA prevents specific interactions but does allow forced Fn-FnBP binding during AFM or stirring in ITC. The bond strength energies calculated from retraction force-distance curves from AFM were orders of magnitude higher than those calculated from desorption data, confirming that a penetrating Fn-coated AFM tip probes multiple adhesins in the outermost cell surface that remain hidden during mild landing of an organism on an Fn-coated substratum, like that during convective diffusional flow.
Population Heterogeneity of
Lactobacillus plantarum
WCFS1 Microcolonies in Response to and Recovery from Acid Stress
Authors: Ingham Colin J. , Beerthuyzen Marke , van Hylckama Vlieg Johan
Published: October 2008

Within an isogenic microbial population in a homogenous environment, individual bacteria can still exhibit differences in phenotype. Phenotypic heterogeneity can facilitate the survival of subpopulations under stress. As the gram-positive bacterium
Lactobacillus plantarum
grows, it acidifies the growth medium to a low pH. We have examined the growth of
L. plantarum
microcolonies after rapid pH downshift (pH 2 to 4), which prevents growth in liquid culture. This acidification was achieved by transferring cells from liquid broth onto a porous ceramic support, placed on a base of low-pH MRS medium solidified using Gelrite. We found a subpopulation of cells that displayed phenotypic heterogeneity and continued to grow at pH 3, which resulted in microcolonies dominated by viable but elongated (filamentous) cells lacking septation, as determined by scanning electron microscopy and staining cell membranes with the lipophilic dye FM4-64. Recovery of pH-stressed cells from these colonies was studied by inoculation onto MRS-Gelrite-covered slides at pH 6.5, and outgrowth was monitored by microscopy. The heterogeneity of the population, calculated from the microcolony areas, decreased with recovery from pH 3 over a period of a few hours. Filamentous cells did not have an advantage in outgrowth during recovery. Specific regions within single filamentous cells were more able to form rapidly dividing cells, i.e., there was heterogeneity even within single recovering cells.
Transposable Element Loads in a Bacterial Symbiont of Weevils Are Extremely Variable
Authors: Dougherty Kevin M. , Plague Gordon R.
Published: October 2008

Not only are transposable elements profuse in the bacterial endosymbiont of maize weevils, but we found that their quantities also vary ∼10-fold among individual weevils. Because multicopy elements can facilitate homologous recombination, this insertion sequence (IS) load variability suggests that these essentially asexual bacteria may exhibit substantial intraspecific genomic variation.