• Title/Summary/Keyword: novel Bacillus

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Improving Protein Production on the Level of Regulation of both Expression and Secretion Pathways in Bacillus subtilis

  • Song, Yafeng;Nikoloff, Jonas M.;Zhan, Dawei
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.963-977
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    • 2015
  • The well-characterized gram-positive bacterium Bacillus subtilis is an outstanding industrial candidate for protein expression owing to its single membrane and high capacity of secretion, simplifying the downstream processing of secretory proteins. During the last few years, there has been continuous progress in the illustration of secretion mechanisms and application of this robust host in various fields of life science, such as enzyme production, feed additives, and food and pharmaceutical industries. Here, we review the developments of Bacillus subtilis as a highly promising expression system illuminating strong chemical- and temperatureinducible and other types of promoters, strategies for ribosome-binding-site utilization, and the novel approach of signal peptide selection. Furthermore, we outline the main steps of the Sec pathway and the relevant elements as well as their interactions. In addition, we introduce the latest discoveries of Tat-related complex structures and functions and the countless applications of this full-folded protein secretion pathway. This review also lists some of the current understandings of ATP-binding cassette transporters. According to the extensive knowledge on the genetic modification strategies and molecular biology of Bacillus subtilis, we propose some suggestions and strategies for improving the yield of intended productions. We expect this to promote striking future developments in the optimization and application of this bacterium.

Anticaries Activity of Antimicrobial Material from Bacillus alkalophilshaggy JY-827

  • Chun, Ju-Yean;Ryu, Il-Hwan;Park, Jung-Sun;Lee, Kap-Sang
    • Journal of Microbiology and Biotechnology
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    • v.12 no.1
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    • pp.18-24
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    • 2002
  • The present study was performed to investigate the excellent microbial anticaries substance, aminoglycoside antibiotic, which is more effective than chlorhexidine for the treatment of dental caries. The aminoglycoside antibiotic against Streptococcus mutans JC-2 from a novel alkaliphilic Bacillus alkalophilshaggy JY-827 exhibited no significant difference at the treatment concentration of $2.5{\times}10^{-7}M$, however, it inhibited the activity of the Streptococcus mutans glucosyltransferase by 70.2% and 99.8% at the concentrations of $2.5{\times}10^{-7}$M\;and\;2.5{\times}10^{-6}M$, respectively. Lineweaver-Burk plot of the inhibitory aminoglycoside antibiotic showed competitive inhibition, with $K_i$ value of $6.4{\times}10^{-6}$ M. The aminoglycoside antibiotic did not show any cytotoxicity against human gingival cells. To evaluate the industrial applicability of the aminoglycoside antibiotic, a toothpaste containing this substance was prepared and tested on the extracted human teeth. The inhibitory rate of tooth calcification and calcium ion elution by the aminoglycoside antibiotic were 50% and 2.5 times, respectively. These results suggested that the aminoglycoside antibiotic from Bacillus alkalophilshaggy JY-827 is an effective agent against dental caries.

Searching Method for New Small RNA in Bacillus subtilis Using Bioinformation (생물정보를 이용하여 바실러스 서브틸리스에서 새로운 Small RNA를 예측하는 방법)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.18 no.1
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    • pp.47-53
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    • 2007
  • In order to find novel sRNA in Bacillus subtilis which would be used to adapt to several conditions, we searched the whole genome of Bacillus subtilis using the following procedure. At first, the locations of recognition sequence of transcription factors such as PerR, OhrR, Fur and Zur were searched in the intergenic region of Bacillus subtilis genome and the locations of rho independent transcription terminator sites were also determined. Based on the information of these locations, the sRNA candidates were chosen by close locations (less than 300 bp) between the recognition site of transcription factors and rho independent transcription terminator site. Than transcription promoter sites were searched in the region of previously identified sRNA candidates and 5 PerR, 1 OhrR, 1 Fur and 1 Zur regulated good sRNA candidates were found.

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Isolation and Characterization of Duck Feather-Degrading Microorganism for Treatment of Recalcitrant Keratinous Waste (난분해성 케라틴 폐기물 처리를 위한 우모 분해 미생물의 분리 및 특성)

  • Go, Tae-Hun;Jeong, Jin-Ha;Lee, Na-Ri;Jeong, Seong-Yun;Park, Geun-Tae;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.21 no.2
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    • pp.253-261
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    • 2012
  • We isolated and characterized novel duck feather-degrading bacteria producing keratinase. Twelve strains were isolated from soil and faces at poultry farm, and decayed feathers. They were identified as Bacillus methylotrophicus, Pseudomonas geniculata, Pseudomonas hibiscicola, Exiquobacterium profundum, Bacillus pumilus, Bacillus amyloliquefaciens, Chryseobacterium indologenes, Bacillus thuringiensis, Thermomonas koreensis, respectively, by phenotypic characters and 16S rRNA gene analysis. Generally, the level of keratinase production was not proportional to feather degradation rate. The highest keratinolytic activity was observed in the culture inoculated with Chryseobacterium indologenes D27. Although all strains did not degrade human hair, strains tested effectively degraded chicken feather(53.8-91.4%), wool(40.4-93.0%) and human nail (51.0-82.9%). These results suggest that strains isolated could be not only used to improve the nutritional value of recalcitrant feather waste but also is a potential candidate for biotechnological processes of keratin hydrolysis.

Distribution and Characterization of Bacillus thuringiensis isolated fro soils in Korea (한국 토양에서 분리된 Bacillus thuringiensis의 분포와 특성조사)

  • 김호산;박현우;이대원;유용만;김정일;강석권
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.344-349
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    • 1995
  • To isolate naturally occurring novel Bacillus thuringiensis strains for useful pest con시 agents, A total of 64 isolates of B. thuringiensis producing spore and crystal were obtained from 583 soil samples in Korea. Toxicity of isolates was assayed against Lepidoptera (Bombyx ori), Diptera (Culex pipens) and Coleoptera (Sitophilus oryzae), respectively. The results showed that B. thuringiensis isolates toxic Lepidoptera are 42.2% , both Lepidoptera and Diptera are 31.3% and Diptera are 20.3%, demonstrating that spectrum of insecticidal activity of B. thuringiensis isolates is various. But no B. thuringiensis isolates have toxicity against Coleoptera.

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Characterization of Bacillus thuringiensis Isolated in Granary Dusts (저곡창고에서 분리된 Bacilus thuringiensis의 특성조사)

  • 김호산;박현우;이대원;유용만;유용만;강석권
    • Korean journal of applied entomology
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    • v.34 no.3
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    • pp.243-248
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    • 1995
  • In order to isolate naturally occurring novel Bacillus thuringiensis strains, we investigated the distribution of B. thuringiensis from granary which exist in Kyong-gi province, Korea. A total of 146 strains of B. thuringiensis producing spore and crystal wre isolated. The toxicity of B. thuringiensis isolates was examined against lepidopteran larvae (Bombyx mori), dipteran larvae (Culex pipiens) and coleopteran larvae (Sitophilus oryzae.), respectively. The results showed that a large number of B. thuringiensis isolates from granary dusts were isolated and most isolates wer toxic to lepidopterous larvae. Also, non-toxic B. thuringiensis isolate was common. In order to isolate many B. thuringiensis, therefore, it suggested that granary is favorable locality.

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A Green Chemical Approach towards the 'One-Pot' Synthesis, Spectral Characterization and in Vitro Antibacterial and Antifungal Activities of Morpholino Pyrimidines (친환경적 One Pot 합성, In Vitro에서 Morpholino Pyrimidines의 분광학적 특성과 항균성 및 항진균성)

  • Kanagarajan, V.;Thanusu, J.;Gopalakrishnan, M.
    • Journal of the Korean Chemical Society
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    • v.53 no.6
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    • pp.731-741
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    • 2009
  • ‘One-pot’ reaction procedure for the synthesis of novel morpholino pyrimidines (10-18) under microwave irradiation in ‘dry media’ in the presence of heterogeneous $NaHSO_4.SiO_2$ catalyst was developed. All the synthesized compounds were screened for their in vitro antibacterial activities against clinically isolated bacterial strains namely Bacillus subtilis, Bacillus cerues, Micrococcus luteus and Salmonella typhii and antifungal activities against fungal strains namely Aspergillus niger, Candida 6 and Candida 51. Structure activity relationship of the synthesized compounds against microbiological results was discussed.

Bioconversion of Tetracycline Antibiotics to Novel Glucoside Derivatives by Single-Vessel Multienzymatic Glycosylation

  • Pandey, Ramesh Prasad;Chu, Luan Luong;Kim, Tae-Su;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.298-304
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    • 2018
  • The single-vessel multienzyme UDP-${\alpha}$-$\text\tiny{D}$-glucose recycling system was coupled with a forward glucosylation reaction to produce novel glucose moiety-conjugated derivatives of different tetracycline antibiotic analogs. Among five tetracycline analogs used for the reaction, four molecules (chlorotetracycline, doxytetracycline, meclotetracycline, and minotetracycline) were accepted by a glycosyltransferase enzyme, YjiC, from Bacillus licheniformis to produce glucoside derivatives. However, the enzyme was unable to conjugate sugar units to rolitetracycline. All glucosides of tetracycline derivatives were characterized by ultraviolet absorbance maxima, ultra-pressure liquid chromatography coupled with photodiode array, and high-resolution quadruple time-of-flight electrospray mass spectrometry analyses. These synthesized glucosides are novel tetracycline derivatives.

Crystal Structure and Biochemical Analysis of a Cytochrome P450 Steroid Hydroxylase (BaCYP106A6) from Bacillus Species

  • Ki-Hwa Kim;Hackwon Do;Chang Woo Lee;Pradeep Subedi;Mieyoung Choi;Yewon Nam;Jun Hyuck Lee;Tae-Jin Oh
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.387-397
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    • 2023
  • Cytochrome P450 (CYP) is a heme-containing enzyme that catalyzes hydroxylation reactions with various substrate molecules. Steroid hydroxylases are particularly useful for effectively introducing hydroxyl groups into a wide range of steroids in the pharmaceutical industry. This study reports a newly identified CYP steroid hydroxylase (BaCYP106A6) from the bacterium Bacillus sp. and characterizes it using an in vitro enzyme assay and structural investigation. Bioconversion assays indicated that BaCYP106A1 catalyzes the hydroxylation of progesterone and androstenedione, whereas no or low conversion was observed with 11β-hydroxysteroids such as cortisol, corticosterone, dexamethasone, and prednisolone. In addition, the crystal structure of BaCYP106A6 was determined at a resolution of 2.8 Å to investigate the configuration of the substrate-binding site and understand substrate preference. This structural characterization and comparison with other bacterial steroid hydroxylase CYPs allowed us to identify a unique Arg295 residue that may serve as the key residue for substrate specificity and regioselectivity in BaCYP106A6. This observation provides valuable background for further protein engineering to design commercially useful CYP steroid hydroxylases with different substrate specificities.