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All Journal Microbiology Indonesia Jurnal Bioteknologi & Biosains Indonesia (JBBI)
Haniyya Haniyya
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Health Professions Immunology & microbiology Medicine & Pharmacology

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The utilization of auto-inducible Plyb promoter and media optimation for cell density-dependent expression of recombinant xylanase in Bacillus subtilis DB104 Haniyya Haniyya; Dini Achnafani; Maria Ulfah; Niknik Nurhayati; Is Helianti
Microbiology Indonesia Vol. 14 No. 1 (2020): March 2020
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1582.039 KB) | DOI: 10.5454/mi.14.1.2

Abstract

Strong promoters are one of the fundamental aspects to increase the level of gene expression, and one of approach to improve the recombinant enzyme productivity so that the efficiency of production cost for enzyme production in industrial scale can be reached. Here we assessed the application of a cell density-dependent promoter and media optimation to promote cell growth and protein expression of Bacillus subtilis without excess usage of inducers. An auto-inducible Pylb promoter that is potential to provide inducer-free enzyme production was cloned and introduced into xylanase recombinant system in B. subtilis DB104 by PCR cloning and protoplast transformation. A 200 bp target gene was successfully inserted in between xynCM1 ORF -coding for B. halodurans CM1 xylanase- and its native promoter sequence at the upstream region. The disruption of the native promoter was intended to replace the native promoter with Pylb. Recombinant xylanase gene under Pylb was successfully expressed in B. subtilis DB104 and the enzyme was produced at stationary phase. Different media with various concentrations of glucose and nitrogen were used to optimize recombinant xylanase expression. It achieved a higher level of xylanase expression compared to wild-type and recombinant xylanase with native promoter B. subtilis in media containing a 2-fold recipe of LB media thus leads to increase cell density and xylanase expression (81.461 U mL-1).
MUTATION OF LIPASE-PRODUCING BACTERIAL ISOLATE FROM PALM OIL EFFLUENT FOR FAT HYDROLYSIS ON POME dadang suhendar; Ika Rahmatul Layly; Gabriela Christy Sabbathini; Deden Rosid Waltam; Edi Wahjono; Catur Sriherwanto; Haniyya Haniyya
Jurnal Bioteknologi dan Biosains Indonesia Vol. 10 No. 1 (2023)
Publisher : BRIN - Badan Riset dan Inovasi Nasional

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Abstract

Lipase is applied to hydrolyzethe residual oil or fat in palm oil mill effluent (POME), which serves as a feedstock for biogas production. A bacterial isolate (Bacillus velezensis) exhibiting lipase activity, previously obtained from Malimping, Pandeglang, Banten, Indonesia, underwentsequential treatments of gamma irradiation (1, 2, and 3 kGy), EMS (ethyl methane sulfonate) mutagenesis (0.5% v/v), and NTG (N-methyl N-nitrosoguanidine) (1 mg mL-1). Following each mutation, lipase activities of the mutant colonies were measured and compared to the wild-type strain. The results revealed that all mutated B. velezensiscolonies exhibited lipase activity values (7.78 ± 0.80 –9.05 ± 0.23 U mL-1) that were not significantly different from the wild-type isolate (8.31 ± 0.01 U mL-1), indicating strong bacterial resistance against the mutagenic treatments. The crude lipase preparation effectively hydrolyzed POME, resulting in a reduction of the COD value (from 131,450 ppm to 88,450 ppm) and O&G content (from 41,400 mg L-1to 5,770 mg L-1) within a 72-hour timeframe.
Co-Authors AA Sudharmawan, AA Catur Sriherwanto Dadang Suhendar deden rosid waltam Dini Achnafani Edi Wahjono Gabriela Christy Sabbathini Ika Rahmatul Layly IS HELIANTI NIKNIK NURHAYATI