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All Journal HAYATI Journal of Biosciences Jurnal Sumberdaya Hayati Majalah Pengabdian Indonesia
Muhamad Azwar Syah
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Decision Sciences, Operations Research & Management Earth & Planetary Sciences Education Immunology & microbiology Industrial & Manufacturing Engineering Languange, Linguistic, Communication & Media Mechanical Engineering Social Sciences

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Cloning and Co-Expression of Lipase and its Specific Foldase from Ralstonia pickettii BK6 Muhamad Azwar Syah; Andreas Adhi Satya; Esti Puspitasari; Antonius Suwanto; Nisa Rachmania Mubarik
HAYATI Journal of Biosciences Vol. 30 No. 1 (2023): January 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.1.71-80

Abstract

This study aimed to obtain a functional lipase (LipRM) from Ralstonia pickettii BK6 through a co-expression involving its foldase. The ORF of the LipRM was 999 bp while the gene encoding of lipase-specific foldase (LifRM) was 1,030 bp. LipRM and LifRM genes were cloned into a plasmid and were successfully co-expressed in Escherichia coli strains to produce functional LipRM. Enzyme activity from partially purified enzymes showed quite surprising results, LipRM activity in E. coli BL21 (DE3) was 25.84 U/ml, while the other strains (DH5α, HB101, S17-1λpir) were 628.98 U/ml, 761 U/ml, and 1206.46 U/ml, respectively. The highest relative activity of LipRM was found at 50-55°C and pH 7-8 with pNP-laurate (C12) as the preferred substrate specificity. LipRM activity was enhanced sharply in the presence of 30% organic solvents (methanol and ethanol) but decreased by more than 50% in the presence of detergents. This study was the first to report heterologous expression of Ralstonia pickettii lipase employing its native foldase resulting in functional lipase from subfamily I.2.
Isolasi dan Karakterisasi Molekuler Gen 16S rRNA Bakteri Lipolitik Asal Limbah Kulit Biji Jambu Mete: Isolation and Molecular Characterization of Lipolytic Bacterial 16S rRNA Gene from Cashew Nutshell Waste Muhamad Azwar Syah
Jurnal Sumberdaya Hayati Vol. 8 No. 1 (2022)
Publisher : Departemen Biologi, Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jsdh.8.1.20-26

Abstract

Lipolytic bacteria attract great attention to various biotechnology industries because of their enzymatic potential. This study aims to isolate and identify lipolytic bacteria from cashew nutshell waste using the 16S rRNA gene as a molecular marker. Lipolytic bacteria were isolated using serial dilutions and inoculated on lipolytic media. A total of 3 isolates of lipolytic bacteria were obtained from cashew nutshell waste based on screening in LA Rhodamine B. The partial sequence of 16S rRNA gene from LB15 amplified using a pair of primers 63F and 1387R having a size of 1238 bp, while BL6 and BK6 were 1283 bp, respectively. Based on genetic distance analysis and phylogenetic reconstruction, we proposed that LB15 be identified as Burkholderia sp. with 99.92% similarity. In addition, because the 16S rRNA gene sequence similarity of BL6 was 99.87% with Paraburkholderia kururiensis strain 979, BL6 was classified as Paraburkholderia kururiensis. Then, isolate BK6 was identified as Ralstonia sp. with a similarity of 99.53%. The similarity value can be used as a reference in determining the identity of bacteria. A bacterium can be categorized as the same species if it has a similarity value of more than 99%.
Training of Microbe Cell Visualization from Fermentation Food at UPT. Integrated Laboratory UHO Muhamad Azwar Syah; Sapto Raharjo; La Ode Muhammad Iman Sulaiman; Jamili; Nasaruddin
Majalah Pengabdian Indonesia Vol. 1 No. 3 (2024): Desember 2024
Publisher : Teras Kampus as a member of PT Palem Edukasi Nusantara

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.69616/maindo.v1i3.236

Abstract

Indonesia has diverse local food commodities that are valuable as a national food source. Microbes serve an important part in food processing, but they can also be harmful and damage food products. The aim of this training is to provide students with skills in understanding microbial cell visualization techniques in food fermentation. Tempeh and the probiotic drink Yakult as sources of fungi and bacteria, respectively, were used in this training. There are four steps for this training consisting of preparation, visualization of mold on tempeh, visualization of bacteria on Yakult as well as training assessment. The findings indicate that students can identify the complex morphology of tempeh mold, which consists of aseptate hyphae, spores, and sporangiophores that sustain the sporangium. Moreover, they exhibited a bacil shape as the morphology of bacteria from Yakult under a light microscope with 400x magnification. Bacterial staining is recommended to visualize bacteria more clearly than without staining. This occurs because bacteria are smaller than mold and have a more translucent hue. The findings of the activity assessment revealed that participants were able to follow the instruction well. This is demonstrated by success indicators in displaying mold morphology, which includes hyphae, sporangiophores, sporangia, and spores. Participants were also able to identify the structure of rod-shaped bacterial organisms, known as bacilli.
Co-Authors Andreas Adhi Satya Antonius Suwanto Esti Puspitasari Jamili La Ode Muhammad Iman Sulaiman Nasaruddin NISA RACHMANIA MUBARIK Sapto Raharjo