Judul: Evaluasi penilaian kud mandiri : Studi kasus pada kud Sinar Jaya dan kud Mekar Jaya
Abstrak: Karya tulis ini bertujuan untuk melihat perkembangan KUD Sinar Jaya dan KUD Mekar Jaya dari tahun kemandiriannya sampai dengan tahun 1993 dan mengevaluasi penilaian KUD.Si­nar Jaya dan KUD Mekar Jaya berdasarkan klasifikasi dan 13 kriteria KUD Mandiri pada tahun buku 1993 serta merumuskan kriteria-kriteria baru yang menggambarkan kemandirian suatu KUD. Penulisan bersumber pada data primer yang dikumpulkan melalui wawancara dengan beberapa anggota KUD, pengurus, pengawas, manajer dan tokoh-tokoh masyarakat serta data se­kunder yang berasal dari laporan RAT, dokumen-dokumen KUD dan literatur yang berkaitan dengan penelitian. KUD Mandiri adalah KUD yang dapat berdiri sendiri dan mampu berswadaya, yaitu bekerja atas dasar kekuatan sendiri dan dapat selalu membuat keputusan-keputusan sendiri yang diperlukan bagi kemajuan ekonomi dan sosial para anggota­nya. Pemerintah (dalam hal ini Departemen Koperasi) telah merumuskan kriteria-kriteria yang dapat menilai kemandirian suatu KUD. Namun ternyata kriteria-kriteria tersebut masih belum sepenuhnya mencirikan kemandirian KUD. Seperti tingkat partisipasi anggota, mutu anggota dan tingkat pengambilan keputusan masih belum tergambarkan dalam 13 kriteria tersebut. Untuk itu perlu dirumuskan kriteria-kriteria baru yang dapat menggambarkan kemandirian KUD. ...
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Judul: Analisis keragaan usaha KUD mandiri dan prospek pengembangan usaha baru (studi kasus di KUD Kediri, Kab. Tabanan-Bali)
Abstrak: Tujuan penelitian ini adalah untuk (1) melihat potensi wilayah kerja dan keragaan usaha yang di lakukan KUD Mandiri, (2) menganalisis perkembangan, pengelolaan modal dan usaha KUD Mandiri, (3) melihat prospek pengembangan usaha baru yang layak di lakukan oleh KUD Mandiri, (4) melihat tingkat partisipasi dan manfaat yang dirasakan sebagai anggota KUD.
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Judul: Extracellular Expression of thermostable Geobacillus stearothermophilus T1.2RQ Lipase for Biodiesel Production
Abstrak: The extracellular expression of enzymes in an excellent secretion host such as Bacillus subtilis is a useful strategy for making enzymes of industrial significance more affordable by lowering the cost of its downstream processing. Lipases are a category of hydrolases that catalyze the hydrolysis of triglycerides to glycerol and free fatty acids. Also, lipases catalyze the hydrolysis and transesterification of other esters as well as the synthesis of esters. They can perform very specific biotransformation reactions and are widely used in food, organic synthesis, detergents, cosmetic, and pharmaceutical industries. Geobacillus stearothermophilus T1.2RQ lipase is an industrially-desired thermostable lipolytic enzyme with an excellent hydrolytic and transesterification activity. This study aimed to express T1.2RQ lipase, in B. subtilis WB800 and ensure its secretion into the extracellular medium. Three signal peptides (AmyQ, Epr, LipA) and two promoters (Pgrac01 and Pgrac100) were used in the secretory-expression of T1.2RQ lipase, based on five constructed expression vectors constructed by restriction cloning and PCR mutagenesis. Sangers sequencing was used to verify constructs to ensure they are in the correct reading frame. Recombinants vectors, pHT43 (plasmid control), pHT43-T1.2RQ, pHT43- T1.2RQ_AmyQ, pHT43-T1.2RQ_Epr, and pHT43-T1.2RQ_LipA, were transformed by electroporation into Bacillus subtilis WB800. The electroporation was successful with the optimum condition of 2500V, 200 ohms, 25µF, and 5milliseconds. Recombinant B. subtilis pHT43 (negative control) and pHT43-T1.2RQlip + signal peptides were streaked on the lipidic agar, LA+TBN. All four recombinants, including the negative control (B. subtilis WB800-pHT43), produced clear zones on the TBN agar plate after overnight incubation (fig 2), indicating the presence of a lipolytic enzyme in all of the recombinant B. subtilis cells. pHT43-T1.2RQlip + signal peptides (AmyQ, Epr, and LipA) have clear zones with significantly higher diameters than B. subtilis WB800-pHT43 (negative control). This phenomenon suggests that the recombinant thermostable lipase T1.2RQ introduced into these cells was successfully expressed, thereby increasing the lipolytic capability of its host cells T1.2RQ lipase was expressed in 50ml terrific broth medium (TB) and the expression induced with 1mM IPTG. Lipase activity assay using p-nitrophenyl esters showed that all three signal peptides directed the secretion of T1.2RQ lipase into the extracellular medium. Signal peptide, LipA, resulted in the highest extracellular activity of 5.6 U/mL, which corresponds to a 6-fold increase over the parent B. subtilis WB800 strain. Meanwhile, signal peptides, AmyQ and Epr produced 4.2 U/mL and 3.4 U/mL activity respectively. SDS-Zymogram analysis confirmed that T1.2RQ lipase was correctly processed and secreted in its original size of 44 kDa. An attempt to further increase lipase expression level by promoter optimization showed that, contrary to expectation, promoter Pgrac100 exhibited similar expression levels as the original Pgrac01. Nevertheless, the demonstration of the successful secretion of T1.2RQ lipase in this work confirms the latter as a promising approach for the industrial production of the enzyme
Keyword: Bacillus subtilis, Expression, Extracellular, Heterologous, Lipase, Thermostable