High yield expression of novel glutaminase free l-asparaginase II of Pectobacterium carotovorum MTCC 1428 in Bacillus subtilis WB800N

2015 ◽  
Vol 38 (11) ◽  
pp. 2271-2284 ◽  
Author(s):  
Sushma Chityala ◽  
Veeranki Venkata Dasu ◽  
Jamal Ahmad ◽  
Reddy Shetty Prakasham
2021 ◽  
Author(s):  
Nagina Rafique ◽  
Saiqa Bashir ◽  
Muhammad Zubair Khan ◽  
Imran Hayat ◽  
Willium Orts ◽  
...  

Pectinolytic enzymes [pectinases] produced by microbes are highly important for their biotechnological use in processing of vegetables and fruits beverages and use in pulp and paper industry. A pectinases, namely endo-polygalacturonase [endo-PGase], encoding gene isolated from Pectobacterium carotovorum, a plant pathogenic strain of bacteria was successfully cloned into a secretion vector pHT43 having σ?-dependent promoter P grac . For enhanced expression analysis, competent cells of Bacillus subtilis (WB800N) were prepared at stationary phase using high salt medium. The recombinant B. subtilis competent cells, harboring the engineered pHT43 with the endo-PGase gene were cultured in 2X-yeast extract tryptone medium. The recombinant endo-PGase enzyme was secreted directly into the medium after 72 hours of the first IPTG induction. The recombinant endo-PGase was screened for its activity at various temperatures and pH ranges. Optimal activity was found at pH 5.0 and a temperature of 40°C with a stability ranging from pH 5.0-9.0. For detection of metal ion effect, recombinant enzyme was incubated with 1mM concentration of; Ca ++ , Mg ++ , Zn ++ , EDTA, K ++ for 45 minutes. Resultantly, Ca ++ , EDTA and Zn ++ strongly inhibited the enzyme activity. The chromatographic analysis of enzymatic hydrolysate of polygalacturonic acid [PGA] and pectin substrates using HPLC and TLC revealed that tri and tetra-galacturonates were the end products of hydrolysis. The study led to the conclusion that endo-PGase gene from the plant pathogenic strain was successfully expressed in Bacillus subtilis and assessed for enzyme production using a very simple medium with IPTG induction. These findings proposed that the Bacillus expression system might be safe for commercial enzyme production as compared to yeast and fungi to escape endotoxins.


2017 ◽  
Vol 117 ◽  
pp. 147-155 ◽  
Author(s):  
Chia-Chi Lin ◽  
Cally Joe San Yap ◽  
Shu-Chen Kan ◽  
Nai-Chi Hsueh ◽  
Liang-Yu Yang ◽  
...  

ACS Omega ◽  
2018 ◽  
Vol 3 (6) ◽  
pp. 7008-7018 ◽  
Author(s):  
Omar E. Tovar-Herrera ◽  
Mabel Rodríguez ◽  
Miguel Olarte-Lozano ◽  
Jimmy Andrés Sampedro-Guerrero ◽  
Adán Guerrero ◽  
...  

2016 ◽  
Vol 23 (3) ◽  
pp. 244
Author(s):  
Hanudin Hanudin ◽  
Abdjad Asih Nawangsih ◽  
Budi Marwoto ◽  
Boedi Tjahjono

<p>Penyakit busuk lunak (PBL) yang disebabkan oleh Pectobacterium carotovorum subsp. carotovorum atau Pseudomonas viridiflava merupakan kendala utama dalam budidaya anggrek. Serangan patogen tersebut sangat merugikan petani, mengingat biaya investasi produksi anggrek tergolong tinggi. Oleh karena itu patogen tersebut harus dikendalikan menggunakan metode pengendalian yang ramah lingkungan, yaitu dengan mengaplikasikan biobakterisida berbahan aktif rizobakteri, seperti Bacillus subtilis dan Pseudomonas fluorescens. Tujuan penelitian ini ialah (1) mendapatkan komposisi bahan aktif dan bahan pembawa biobakterisida yang efektif mengendalikan penyakit busuk lunak pada anggrek Phalaenopsis, (2) mengetahui perubahan reaksi kimia formula biobakterisida dan pertumbuhan populasi bahan aktif (rizobakteri B. subtilis dan P. fluorescens) pada kondisi sebelum dan setelah difermentasikan, dan (3) mengetahui kompatibilitas antara B. subtilis, P. fluorescens, dan bahan pembawa biobakterisida. Percobaan dilaksanakan mulai Bulan Mei sampai dengan Desember 2009 di Laboratorium Bakteriologi Departemen Proteksi Tanaman, Institut Pertanian Bogor, dan Laboratorium Bakteriologi serta Rumah Kaca, Balai Penelitian Tanaman Hias, di Segunung, Cianjur, Jawa Barat. Ruang lingkup penelitian meliputi pembuatan propagul rizobakteri sebagai bahan aktif biobakterisida, pembuatan formula biobakterisida, uji viabilitas bahan aktif, dan uji kemangkusan biobakterisida pada tanaman anggrek di rumah kaca. Hasil penelitian menunjukkan bahwa (1) perlakuan gabungan antara B. subtilis B12 dan P. fluorescens Pf10 yang difermentasikan dalam media ekstrak kotoran cacing (kascing) dan molase, merupakan perlakuan yang konsisten dapat menekan PBL pada anggrek Phalaenopsis dengan persentase penekanan sebesar 80%, (2) reaksi kimia formula biopestisida pada kondisi sebelum dan setelah fermentasi diindikasikan dengan perubahan pH basal medium yang sebelum fermentasi menunjukkan pH 3,75 dan berubah menjadi pH 3,50 setelah difermentasikan. Pertumbuhan populasi mikrob antagonis setelah fermentasi meningkat secara signifikan bila dibandingkan pada kondisi sebelum difermentasikan, dan (3) isolat bahan aktif (B. subtilis dan P. fluorescens) bersifat kompatibel dengan bahan pembawanya (ekstrak kascing dan molase).</p>


2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Dhouha Ghribi ◽  
Semia Ellouze-Chaabouni

Interest in biosurfactants has increased considerably in recent years, as they are potentially used in many commercial applications in petroleum, pharmaceuticals, biomedical, and food processing industries. Since improvement of their production was of great importance to reduce the final coast, cultural conditions were analyzed to optimize biosurfactants production from Bacillus subtilis SPB1 strain. A high yield of biosurfactants was obtained from a culture of B. subtilis using carbohydrate substrate as a carbon source; among carbohydrates, glucose enhanced the best surfactin production. The optimum glucose concentration was 40 g/L. Higher amount of biosurfactants was obtained using 5 g/L of urea as organic nitrogen source and applying C/N ratio of 7 with ammonium chloride as inorganic nitrogen source. The highest amount of biosurfactants was recorded with the addition of 2% kerosene. Moreover, it was shown, using an automated full-controlled 2.6 L fermenter, that aeration of the medium, which affected strongly the growth regulated biosurfactants synthesis by the producing cell. So that, low or high aerations lead to a decrease of biosurfactants synthesis yields. It was found that when using dissolved oxygen saturation of the medium at 30%, biosurfactants production reached 4.92 g/L.


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