A Bacillus subtilis Strain ZJ20 with AFB1 Detoxification Ability: A Comprehensive Analysis
Meixue HuangGuo JingYanyan JiaChengshui LiaoLei HeJing LiYing WeiSongbiao ChenJian ChenKe ShangRongxian GuoKe DingZuhua Yu
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As a class I carcinogen, aflatoxin can cause serious damage to various tissues and organs through oxidative stress injuries. The liver, as the target organ of AFB1, is the most seriously damaged. Biological methods are commonly used to degrade AFB1. In our study, the aflatoxin B1-degrading strain ZJ20 was screened from AFB1-contaminated feed and soil, and the degradation of AFB1 by ZJ20 was investigated. The whole genome of strain ZJ20 was analyzed, revealing the genomic complexity of strain ZJ20. The 16S rRNA analysis of strain ZJ20 showed 100% identity to Bacillus subtilis IAM 12118. Through whole gene functional annotation, it was determined that ZJ20 has high antioxidant activity and enzymatic activity; more than 100 CAZymes and 11 gene clusters are involved in the production of secondary metabolites with antimicrobial properties. In addition, B. subtilis ZJ20 was predicted to contain a cluster of genes encoding AFB1-degrading enzymes, including chitinase, laccase, lactonase, and manganese oxidase. The comprehensive analysis of B. subtilis provides a theoretical basis for the subsequent development of the biological functions of ZJ20 and the combinatorial enzyme degradation of AFB1.Keywords:
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A biocontrol strain,XF-1,was identified as Bacillus subtilis based on morphology,physiology,biochemistry and biotechnology,which could control clubroot of cruciferous crops effectively.The sequence of 16S rRNA gene has 99% identity to B.subtilis according to the amplified fragment of 1513 bp with a pair of universal primers,P0 and P6,of bacteria.On co-culture plates this strain showed high suppression effect on 21 fungal pathogens involved in oomycetes,ascomycetes,basidiomycetes and deuteromycetes.
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Most strains with capability of producing 2,3-butanediol(2,3-BDO) from glucose were isolated from different natural environment samples.The Strain 6-7 with high capability of producing 2,3-BDO was screened from the isolated strains and the production of 2,3-BDO reached 49.6 g/L.In addition to general morphological and biochemical characteristics,the strain 6-7 was identified by 16S rDNA sequence and systematic analysis.The results showed that 16S rDNA sequence of strain 6-7 had similarity of 99% with Bacillus subtilis strain BIHB332,suggesting that the strain 6-7 is one of Bacillus subtilis species.It is an environmentally friendly strain and Bacillus subtilis is generally regarded as safe microorganism used for industrial production.This strain showed the potential application in the future.
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A Tween-80-degrading novel marine Bacillus strain, N10, has recently been isolated in Alexandria University, Egypt. The taxonomic position of this endospore forming bacterium was investigated on the basis of fatty acid analysis and 16S rRNA gene sequencing. Comparative computer database analyses revealed that the bacterium is a Bacillus subtilis strain. The gene encoding the small acid-soluble protein γ-type (SASP-B), sspE, was successfully utilized in this study as a tool for discrimination between the two B. subtilis subspecies W23 and 168. Based on the alignment of 16S rRNA sequences and analysis of SASP-B relatedness, it has been demonstrated that the novel marine B. subtilis strain N10 is more closely related to the B. subtilis reference strain W23 than to 168. The strain, N10, has been deposited in the Bacillus Genetic Stock Center (BGSC) and assigned the accession number 3A17.
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In order to obtain the thermostable desizing enzyme,which can be used in textile industry,a strain which can excrete thermostable α-amylase was isolated from soil,and the characteristics of its crude enzyme were studied.The results show that the strain is identified preliminarily as Bacillus subtilis,temporarily named Bacillus subtilis C1.In the basal medium,the strain C1 reaches its stationary phase within 24 h at pH 7 and 45 ℃.And at 68 h,it reaches the highest enzyme activity of 185.6 U/mL.The optimum pH value is 6 and the optimum temperature is 70℃.This enzyme is stable to high temperature,and 81 % of enzyme activity is retained after being disposed at 70 ℃ for 1 h.
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Purpose To isolate and to identify strains producing fibrinolytic enzyme.Methods Strains producing fibrinolytic enzyme were isolated by LB-fibrin plate at first and secondly by shaking flask fermentation.Morphological,physiological and biochemical characteristics and 16S rDNA were used to identify strain.Results Five strains with high fibrinolytic activity were screened from soil and the results indicated that the BS-26 strain had the highest fibrinolytic activity,and its characters were very similar to Bacillus subtilis.A phylogenetic tree was constructed by comparing with the validly published 16S rDNA sequences of the related strains from GenBank using the Neighbor-Joining method.The 16S rDNA sequence of strain BS-26 shared 99.63% similarity and probability with Bacillus subtilis.Conclusion BS-26 strain was identified as Bacillus subtilis.
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Extraction and characterization of extracellular chitinase from Bacillus subtilis B 298 have been done. Growth curve determination of B. subtilis B 298, production curve determination of crude extract chitinase from B. subtilis B 298, and partial biochemical characterization of crude extract chitinase have been achieved in this study. Optimum growth of B. subtilis B 298 was achieved at logarithmic phase within 9 hours incubation time, so it was used as inoculum for enzyme production. According to production curve of the enzyme, it was known that incubation time which gave the highest chitinase activity of 15 hours with activity of 6.937 U/mL respectively. Effect of various temperatures on chitinase activity showed that optimum activity was achieved at 40°C with an activity of 5.764 U/mL respectively. Meanwhile, the optimum pH for chitinase activity was achieved at pH of 5.0 with an activity of 6.813 U/mL respectively. This enzyme was then classified as metalloenzyme due to the decline of the activity by EDTA addition. All divalent cations tested acted as inhibitors.
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