Optimization and characterization of the depolymerase gene from Fusarium perfringens Expression in Pichia pastoris
Abstract
poly (butylene succinate) is a biodegradable plastic that can replace traditional plastics, which is helpful to solve the problem of white pollution, the key to the rapid and effective degradation of microorganisms or depolymerization Enzymes.The recombinant Pichia pastoris expression system was constructed to realize (FSC)EFFICIENT EXPRESSION.Codon Optimization Based on Pichia pastoris preferenceFSCGene codons of Pichia PastorisX33Achieving its recombinant expression in.The fermentation conditions of the strain were optimized by single factor experiments..The results show that the optimizedFSCIn gene sequence157One base changed,G + CContent59.6%Drop48.3%, Sequence homology is77.34%;Constructed recombinant expression vectorPpiczAlpha- FSCTransferred to Pichia PastorisX33, Combined with resistance plate screening,SDS-PAGEAndWestern BoltAnd the enzyme activity of shake flask fermentation was determined.1.Recombinant strain with high enzyme yieldL1;Further determine its fermentation conditions:Medium InitiationPH 6.0, Shaker speed220 r/minMethanol supplement1%, Inoculation amount8%Training time72 h, Incubation temperature30Under the optimized conditions, the enzyme activity of fermentation broth was up110 U/ml.
Keywords
Full Text:
PDFReferences
RIchardKalra B.Biodegradable Polymers. environ-ment[J].Science2002297:803-807.
Shah AAHasan FHameedEt al.Biological degradation. Plastics:A comprehensive review[J].Biotechnology ad-vances,2008,26:246-265.
Maeda H,Yamagata y,Abe K,Et al.Purification and charac-characterization of a biodegraded plas-tic-upgrading enzyme from as-Aspergillus Oryzae[J].Applied Microbiology Biotechnology,2005,67:778-788.
Uchida H,Shigeno-akutsu y,Nomura n,Et al.Cloning and Sequence Analysis of poly(Tetramethylene Succinate)Poly-merase from Acidovorax delafieldii strain BS-3[J].Journal of bioscience and bioengineering,2002,93(2.):245-247.
Kunioka m,Ninomiya F,Funabashi m.Biodegrayment OF POLY Butylene Succinate)Powder in a controlled post at 58 Evaluated by naturally-occurring carbon 14 amounts in evolved Co2.Based on the ISO 14855-2 Method[J].Mobile sci- Ences,2009,10(10):4267-4283.
Akutsu-shigeno y,Teeraphatpornchai t,Teamtisong K,Et al.Cloning, sequencing. a Poly(DL-Lactic Acid)Depoly-merase gene from Paenibacillus amylolyticus strain TB-13, its functional expression. Escherichia coli[J].Applied, environmental Microbiology200369(5):2498-2504.
Masaki KKamini. NRIkeda HEt al.Cutinase-like enzyme. yeast Cryptococcus sp.Strain S-2 hydrolyzes Polylactic acid, other biodegradable plastics[J].Applied, Environ-mental Microbiology200571(11):7548-7550.
Shinozaki YMorita TCao XHEt al.Biodegradable plastic-degrading enzyme from pseudozyma Antarctica:CloningSe-quencing, Characterization[J].Applied Microbiology Bio-technology201397(7/):2951-2959.
Mao HLiu HGao twig u & ZEt al.Biodegradation. poly(Butylene Succinate)By Fusarium sp.FS1301, purification, char-acterization. poly(Butylene Succinate)Depolymerase[J].Polymer Deg-radation, stability2015114:1-7/.
Wu Jie chang xiao xue more than River and., Red yeast engineering bacteria high-density fermentation research Research and progress [J]., China biological engineering magazine,201636(1):108-114.
King zhan yong.Poly (lactic acid) degradation bacteria of breeding and Its depolymerase study [D].Changchun:Northeast Normal University,2011.
Strict into Yin Li Feng weeks Tami-Lynn and.Alkalophilic bacillus of heat-resistant alkali-resistant Endo xylanase gene of codon optimization and in, Red Yeast in expression [J].Guangxi Science,201724(1):100-105.
Zhang Yan Li Meng wax Zhang Jianguo.γ-Cyclodextrin glycosylation transfer enzyme in Pichia pastoris Yeast in Efficient expression [J].Industrial microbial,201747(1):24-30.
Bed Bo Yun white renhui King chun di and.Thermophilic cotton bacteria wood glucosidaseXyl43Gene optimization and Its in, Red Yeast in efficient expression [J].Microbiology Bulletin,201744(1):9-19.
King Kin Wing Liu Dany summer rain and.Optimization codon and induced by temperature improve white Rhizopus lipase in Pichia pastoris Yeast in expression [J].Food and fermentation Industrial
Refbacks
- There are currently no refbacks.
