## 3 Results and Discussion  
### Alakalophilic Pectinase-Producing Bacteria  
The pure cultured alkali-tolerant bacterial colonies APPB1 and APPB2 were isolated from wet paddy land soil and kenaf retting niche. Both isolates were able to grow on an adapted alkaline (pH 9.0) nutrient agar medium. Screening by the plate assay methodrevealed that the cultured bacteria were able to grow in alkaline in pH media with 1% Na2 CO3 solution. According to [41], bacteria and other microbes that are optimally grown at pH 9.0 or above are considered alkalophilic bacteria. These isolates did not necessarily require obligate alkaliphic; however, elucidation at multiple pH conditions showed optimal growth at an elevated pH (-9). Once they grew on H1 media, a lot of colonies were visible on the plates and countable numbers of colonies (30-300 cfu/mL) were observed in the diluted samples. Alkalophilic bacteria can survive in harsh environmental conditions and maintain cellular homeostasis due to their proton-motive force for ATP synthesis [42]. Along with extreme alkaline niches, alkaliphic bacteria are abundant in elevated pH niches, such as serpentinizing sites and wet paddy soil, where continuous bio-recycling occurred. However, the core task in advancing the retting process is to obtain strains capable of producing pectinase enzymes. The pectinolytic potency of the isolates was confirmed by their pectin hydrolysis potency in a solid pectin agar medium (Figure 1). The screening results show that both isolates were able to hydrolyze pectin; therefore, these isolates were designated as alkalophilic pectinase-producing bacteria (APPB). Even though multiple enzymes are required to degrade heterogeneous cell wall components, pectinase is the main retting enzyme determining the retting magnitude, and applications of high efficiency pectinolytic bacterial strains are advantageous in decreasing the retting period, thus enhancing fiber quality [25, 43]. The quantitative multiple enzymatic activities of these isolates are presented in Figure 2. The majority of naturally occurring microorganisms do not produce enzymes in adequate levels for industrial use, and many of them lack the desired characteristics for applications. Hence, both the stains showed their enzymatic potency to be used as retting agents and are comparable with other reported results [25, 44]. In this paper, the xylano-pectinolytic activity of both isolates was associated with cellulase activity (Figure 2). The xylano-pectinolytic bacterial strains deprived of cellulase activities were considered critical for the degumming process. On the contrary, [PERSON] et al. [13] stated that, in addition to pectinases, hemicellulaes and cellulases are also required to obtain good retting results. In general, cellulase are not preferred for degumming as those preferentially attack the fiber nodes or kink bands and weaken the fibers. Even so, microbial pectinases are well-mixed with cellulases and hemicellulaes activities [15, 45]. The bacterial extracellular enzymatic activity was considered the critical biocatalyst in the degradation and utilization of organic polymers. In this paper, the enzymatic activities of the isolates were estimated using the extracellular crude enzymes of all three enzymatic activities, which is consistent with the findings of other studies, when compared with the crude supernatant enzyme assay. The findings of the crude enzymatic activities of both isolates are consistent with the results of previous studies, such as those reported by [25, 44, 46] for _Bacillus_ spp. Figure 1: Formation of a pectin hydrolysis zone by alkalophilic pectinase-producing bacteria, APPB1 (**b**) and APPB2 (**c**), compared with a no hydrolysis zone (**a**). Both isolates were Gram-positive, rod-shaped bacilli, spore-forming, and able to grow in facultative anaerobic conditions, with differences in morphology and biochemical features. The colony of APPB1 had a marked yellowish color with slender cells, while APPB2 had a marked pale yellowish color with a robust cell structure (Figure 3).