常压室温等离子体诱变选育木聚糖酶高产菌及其酶学性质研究
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摘要
利用常压室温等离子体(ARTP)技术对黑曲霉(Aspergillus niger)FXY进行诱变处理,并对菌株的遗传稳定性及所产木聚糖酶的酶学性质进行了研究。结果表明,筛得一株高产木聚糖酶且能稳定遗传的菌株ARTP-82,其所产木聚糖酶酶活为175.33 U/m L,较出发菌株提高了120.8%。酶活的提高与酶比活力增加有关、与菌株生物量增加无关;该木聚糖酶的最适温度为45.0℃、最适p H值为7.0,在该温度和pH值条件下酶的稳定性良好。
The Aspergillus niger FXY was mutated by atmospheric and room temperature plasma(ARTP), and the hereditary stability of the strain and the enzymatic properties of the xylanase were studied. The results showed that a strain ARTP-82 with high yield xylanase and stable hereditary was screened, and the xylanase activity was 175.33 U/ml, which was 120.8% higher than that of the original strain. The reason for the increase in xylanase activity was related to the increase of enzyme specific activity, instead of the increase of strain biomass. The optimum temperature and pH of xylanase were 45.0 ℃ and 7.0, respectively, and the stability of the enzyme under the condition was good.
The Aspergillus niger FXY was mutated by atmospheric and room temperature plasma(ARTP), and the hereditary stability of the strain and the enzymatic properties of the xylanase were studied. The results showed that a strain ARTP-82 with high yield xylanase and stable hereditary was screened, and the xylanase activity was 175.33 U/ml, which was 120.8% higher than that of the original strain. The reason for the increase in xylanase activity was related to the increase of enzyme specific activity, instead of the increase of strain biomass. The optimum temperature and pH of xylanase were 45.0 ℃ and 7.0, respectively, and the stability of the enzyme under the condition was good.
引文
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[2]LIU X,LIU Y,JIANG Z,et al.Biochemical characterization of a novel xylanase from Paenibacillus barengoltzii and its application in xylooligosaccharides production from corncobs[J].Food Chem,2018,264(10):310-318.
[3]张燕青,张超群,王浩猛.木聚糖酶的分子改造方法及其工业应用研究现状[J].中国酿造,2018,37(1):25-29.
[4]PARAB P,KHANDEPARKER R,AMBERKAR U,et al.Enzymatic saccharification of seaweeds into fermentable sugars by xylanase from marine Bacillus sp.strain BT21[J].Biotech,2017,7(5):296.
[5]DHIMAN S S,GARG G,SHARMA J,et al.Characterization of statistically produced xylanase for enrichment of fruit juice clarification process[J].New Biotechnol,2011,28(6):746-755.
[6]KHAMBHATY Y,AKSHAYA R,RAMA S C,et al.A logical and sustainable approach towards bamboo pulp bleaching using xylanase from Aspergillus nidulans[J].Int J Biol Macromol,2018,118(A):452-459.
[7]陈洪洋,蔡俊,林建国,等.木聚糖酶的研究进展[J].中国酿造,2016,35(11):1-6.
[8]WANG J,MAI G,LIU G,et al.Molecular cloning and heterologous expression of an acid-stable endoxylanase gene from Penicillium oxalicum in Trichoderma reesei[J].J Microbiol Biotechn,2013,23(2):251-259.
[9]CANAKCI S,CWVHER Z,INAN K,et al.Cloning,purification and characterization of an alkali-stable endoxylanase from Thermophilic Geobacillus sp.71[J].World J Microb Biot,2012,28(5):1981-1988.
[10]POLIZELI M L,RIZZATTI A C,MONTI R,et al.Xylanases from fungi:properties and industrial applications[J].Appl Microbiol Biot,2005,67(5):577-591.
[11]郑丽丽,盛占武,韩冰莹,等.不同诱变方法对黑曲霉产木聚糖酶能力的影响[J].生物技术通报,2013(12):146-150.
[12]KUMAR V,CHHABRA D,SHUKLA P.Xylanase production from Thermomyces lanuginosus VAPS-24 using low cost agro-industrial residues via hybrid optimization tools and its potential use for saccharification[J].Bioresource Technol,2017,243:1009-1019.
[13]吴仁智,黄俊,芦志龙,等.酸性木聚糖酶产生菌XYW5的筛选及酶学性质[J].广西科学,2018,25(6):669-677.
[14]LI D,LI X,DANG W,et al.Characterization and application of an acidophilic and thermostableβ-glucosidase from Thermofilum pendens[J].J Biosci Bioeng,2013,115(5):490-496.
[15]DONG T T,GONG J S,GU B C,et al.Significantly enhanced substrate tolerance of Pseudomonas putida nitrilase via atmospheric and room temperature plasma and cell immobilization[J].Bioresource Technol,2017,244(Pt 1):1104-1110.
[16]CHENG G,XU J,XIA X,et al.Breeding L-arginine-producing strains by a novel mutagenesis method:Atmospheric and room temperature plasma(ARTP)[J].Preparat Biochem,2016,46(5):509-516.
[17]ZHANG X,ZHANG X F,LI H P,et al.Atmospheric and room temperature plasma(ARTP)as a new powerful mutagenesis tool[J].Appl Microbiol Biotechn,2014,98(12):5387-5396.
[18]MESHRAM M,KULKARNI A,JAYARAMAN V K,et al.Optimal xylanase production using Penicilium janthinellum NCIM 1169:A model based approach[J].Biochem Eng J,2008,40(2):348-356.
[19]李娟,张耀庭,曾伟,等.应用考马斯亮蓝法测定总蛋白含量[J].中国生物制品学杂志,2000,13(2):118-120.
[20]NING Z,LONG Y.Effect of N+ion implantation on antioxidase activity in Blakeslea trispora[J].Radiat Phys Chem,2008,77(9):1046-1049.
[21]林良斌.SOS修复和突变生产[J].湖南科技大学学报(社会科学版),1991,12(3):92-97.
[22]白冠军,任衍钢,宋玉奇,等.紫外线诱导DNA损伤及修复研究的起因与发展[J].生物学通报,2017,52(3):58-62.
[23]CAO S,ZHOU X,JIN W,et al.Improving of lipid productivity of the oleaginous microalgae Chlorella pyrenoidosa via atmospheric and room temperature plasm(ARTP)[J].Bioresource Technol,2017,244(Pt 2):1400-1406.