Significantly enhancing recombinant alkaline amylase production in Bacillus subtilis by integration of a novel mutagenesis-screening strategy with systems-level fermentation optimization View Full Text


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Article Info

DATE

2016-12

AUTHORS

Yingfang Ma, Wei Shen, Xianzhong Chen, Long Liu, Zhemin Zhou, Fei Xu, Haiquan Yang

ABSTRACT

BACKGROUND: Alkaline amylase has significant potential for applications in the textile, paper and detergent industries, however, low yield of which cannot meet the requirement of industrial application. In this work, a novel ARTP mutagenesis-screening method and fermentation optimization strategies were used to significantly improve the expression level of recombinant alkaline amylase in B. subtilis 168. RESULTS: The activity of alkaline amylase in mutant B. subtilis 168 mut-16# strain was 1.34-fold greater than that in the wild-type, and the highest specific production rate was improved from 1.31 U/(mg·h) in the wild-type strain to 1.57 U/(mg·h) in the mutant strain. Meanwhile, the growth of B. subtilis was significantly enhanced by ARTP mutagenesis. When the agitation speed was 550 rpm, the highest activity of recombinant alkaline amylase was 1.16- and 1.25-fold of the activities at 450 and 650 rpm, respectively. When the concentration of soluble starch and soy peptone in the initial fermentation medium was doubled, alkaline amylase activity was increased 1.29-fold. Feeding hydrolyzed starch and soy peptone mixture or glucose significantly improved cell growth, but inhibited the alkaline amylase production in B. subtilis 168 mut-16#. The highest alkaline amylase activity by feeding hydrolyzed starch reached 591.4 U/mL, which was 1.51-fold the activity by feeding hydrolyzed starch and soy peptone mixture. Single pulse feeding-based batch feeding at 10 h favored the production of alkaline amylase in B. subtilis 168 mut-16#. CONCLUSION: The results indicated that this novel ARTP mutagenesis-screening method could significantly improve the yield of recombinant proteins in B. subtilis. Meanwhile, fermentation optimization strategies efficiently promoted expression of recombinant alkaline amylase in B. subtilis 168 mut-16#. These findings have great potential for facilitating the industrial-scale production of alkaline amylase and other enzymes, using B. subtilis cultures as microbial cell factories. More... »

PAGES

13

References to SciGraph publications

  • 2015-07. Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2014-06. Atmospheric and room temperature plasma (ARTP) as a new powerful mutagenesis tool in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2011-09. Production of nattokinase by high cell density fed-batch culture of Bacillus subtilis in BIOPROCESS AND BIOSYSTEMS ENGINEERING
  • 2008-10. An effective and simplified pH-stat control strategy for the industrial fermentation of vitamin B12 by Pseudomonas denitrificans in BIOPROCESS AND BIOSYSTEMS ENGINEERING
  • 2007-09. Optimization of riboflavin production by recombinant Bacillus subtilis RH44 using statistical designs in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2004-12. Enhanced amylase production by Bacillus subtilis using a dual exponential feeding strategy in BIOPROCESS AND BIOSYSTEMS ENGINEERING
  • 2016-12. Homogeneity and heterogeneity in amylase production by Bacillus subtilis under different growth conditions in MICROBIAL CELL FACTORIES
  • 2010-04. Analysis of Carbon Metabolism and Improvement of γ-Polyglutamic Acid Production from Bacillus subtilis NX-2 in APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
  • 2013-12. Cloning and enhancing production of a detergent- and organic-solvent-resistant nattokinase from Bacillus subtilis VTCC-DVN-12-01 by using an eight-protease-gene-deficient Bacillus subtilis WB800 in MICROBIAL CELL FACTORIES
  • 2015-01. An exceptionally cold-adapted alpha-amylase from a metagenomic library of a cold and alkaline environment in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2016-12. Development of an efficient autoinducible expression system by promoter engineering in Bacillus subtilis in MICROBIAL CELL FACTORIES
  • Journal

    TITLE

    Journal of Biological Engineering

    ISSUE

    1

    VOLUME

    10

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s13036-016-0035-2

    DOI

    http://dx.doi.org/10.1186/s13036-016-0035-2

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1038196793

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/27777616


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