Breeding Aspergillus oryzae with high-productive protease and its influence on soy sauce fermentation

  • FAN Jiaxun ,
  • LIU Song ,
  • LU Xinyao ,
  • CHEN Jian
Expand
  • 1(National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China)
    2(School of Biotechnology, Jiangnan University, Wuxi 214122, China)

Received date: 2021-02-09

  Revised date: 2021-03-31

  Online published: 2021-11-30

Abstract

In order to improve the breeding efficiency of Aspergillus oryzae, a high-throughput screening method based on flow cytometry was constructed. After four rounds of mutation using atmospheric and room temperature plasma, the strain H34 with 145.6% increase of protease activity was screened. The increased protease activities in strain H34 were mainly aspergillopepsin-1, alkaline protease, and neutral protease 2 homolog AO090001000135. The soy sauce produced by H34 strain was superior in total nitrogen, amino acid nitrogen, organic acid, amino acid, and flavor substance. The results provide an excellent strain for soy sauce fermentation and important methodological reference for mutation breeding of other filamentous fungi.

Cite this article

FAN Jiaxun , LIU Song , LU Xinyao , CHEN Jian . Breeding Aspergillus oryzae with high-productive protease and its influence on soy sauce fermentation[J]. Food and Fermentation Industries, 2021 , 47(21) : 1 -8 . DOI: 10.13995/j.cnki.11-1802/ts.027031

References

[1] DEVANTHI P V P,GKATZIONIS K.Soy sauce fermentation:Microorganisms,aroma formation,and process modification[J].Food Research International,2019,120:364-374.
[2] XU Y,WANG D,FAN W L,et al.Traditional Chinese biotechnology[J].Advances in Biochemical Engineering/Biotechnology,2010,122:189-233.
[3] 潘力, 李立风,彭昶,等.酱油曲霉孢子原生质体的制备与紫外诱变育种[J].食品与发酵工业,2006,32(8):1-4.
PAN L,LI L F,PENG C,et al.Protoplast preparation and regeneration of Aspergillus sojae spore and improvement of its protease activity by UV treatment[J].Food and Fermentation Industries,2006,32(8):1-4.
[4] ZHAO G Z,HOU L H,LU M F,et al.Construction of the mutant strain in Aspergillus oryzae 3.042 for abundant proteinase production by the N+ ion implantation mutagenesis[J].International Journal of Food Science & Technology,2012,47(3):504-510.
[5] XU D F,PAN L,ZHAO H F,et al.Breeding and identification of novel koji molds with high activity of acid protease by genome recombination between Aspergillus oryzae and Aspergillus niger[J].Journal of Industrial Microbiology & Biotechnology,2011,38(9):1 255-1 265.
[6] 舒冬梅, 王德良,宋绪磊,等.常压室温等离子体诱变(ARTP)及高通量筛选高产蛋白酶米曲霉的初探[J].中国调味品,2016,41(12):67-73.
SHU D M,WANG D L,SONG X L,et al.Initial study on high proteinase-producing Aspergillus oryzae with ARTP mutation and high-throughput screening[J].China Condiment,2016,41(12):67-73.
[7] ADAN A,ALIZADA G,KIRAZ Y,et al.Flow cytometry:Basic principles and applications[J].Critical Reviews in Biotechnology,2017,37(2):163-176.
[8] OTTENHEIM C,NAWRATH M,WU J C.Microbial mutagenesis by atmospheric and room-temperature plasma (ARTP):The latest development[J].Bioresources & Bioprocessing,2018,5(1):1-14.
[9] 柴阿丽, 韩云,武军,等.基于FDA-PI双荧光复染法的茄病镰刀菌孢子活性检测[J].中国农业科学,2015,48(14):2 757-2 766.
CHAI A L,HAN Y,WU J,et al.Determination of spore viability of Fusarium solani based on dual fluorescence assay[J].Scientia Agricultura Sinica,2015,48(14):2 757-2 766.
[10] 朱金勇, 黄河清.MALDI-TOF质谱技术分析细胞与组织多肽组成的研究进展[J].分析仪器,2004(2):5-9.
ZHU J Y,HUANG H Q.Progress of research on analysis of peptide components in cell and tissue with MALDI-TOF mass spectrometry[J].Analytical Instrumentation,2004(2):5-9.
[11] ODA K,KAKIZONO D,YAMADA O,et al.Proteomic analysis of extracellular proteins from Aspergillus oryzae grown under submerged and solid-state culture conditions[J].Applied and Environmental Microbiology,2006,72(5):3 448-3 457.
[12] 房峻, 刘佳乐,彭玉慧,等.强化嗜盐四联球菌对模拟低盐酱油发酵的影响[J].中国酿造,2020,39(12):146-152.
FANG J,LIU J L,PENG Y H,et al.Effect of enrichment of Tetragenococcus halophilus on simulated fermentation of low-salt soy sauce[J].China Brewing,2020,39(12):146-152.
[13] 朱莉, 许长华.酱油关键风味物质及其功能与发酵工艺研究进展[J].食品与发酵工业,2018,44(6):287-292.
ZHU L,XU C H.Research advances on the key flavor substance of soy sauce and its functions and fermentation technology[J].Food and Fermentation Industries,2018,44 (6):287-292.
[14] 梁姚顺, 朱新贵,孙胜枚,等.几种含氮物质对酱油氨基酸态氮及全氮测定值的影响[J].中国调味品,2011,36(10):76-78.
LIANG Y S,ZHU X G,SUN S M,et al.Study on the effect of some nitrogenous compounds on the determination of amino acid nitrogen and total nitrogen in soy sauce[J].China Condiment,2011,36 (10):76-78.
[15] 陈智慧, 史梅,王秋香,等.用凯氏定氮法测定食品中的蛋白质含量.首届中国兽药大会动物药品学暨中国畜牧兽医学会动物药品学分会2008学术年会论文集[C].天津:中国动物保健品协会,2008.
CHEN Z H,SHI M,WANG Q X,et al.Determination of protein content in food by Kjeldahl nitrogen determination method.Proceedings of the 1 st Chinese Veterinary Drug Congress and the 2008 Academic Annual Meeting of the Chinese Society of Animal Science and Veterinary Medicine[C].Tianjin:China Animal Health Products Association, 2008.
[16] 仇钰莹. 浓香型白酒中氨基甲酸乙酯形成途径解析[D].无锡:江南大学,2016.
QIU Y Y.Analysis of ethyl carbamate formation during Luzhou-flavor spirits fermentation process[D].Wuxi:Jiangnan University,2016.
[17] 王银辉, 沈小梅,马雷,等.超高效液相色谱:三重四级杆串联质谱仪(UPLC-MS/MS)同时测定白酒中7种有机酸的分析研究[J].酿酒,2016,43(1):43-46.
WANG Y H,SHEN X M,MA L,et al.Ultra performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-MS/MS) simultaneously determine 7 kinds of organic acids in liquor[J].Liquor Making,2016,43(1):43-46.
[18] 童佳, 赵国忠,赵建新,等.米曲霉发酵高盐稀态酱油过程中典型挥发性风味物质的形成[J].中国酿造,2017,36(5):22-29.
TONG J,ZHAO G Z,ZHAO J X,et al.Formation of typical volatile flavor compounds in soy sauce with high salt liquid state fermentation by Aspergillus oryzae[J].China Brewing,2017,36(5):22-29.
[19] ROTMAN B,PAPERMASTER B W.Membrane properties of living mammalian cells as studied by enzymatic hydrolysis of fluorogenic esters[J].Proceedings of the National Academy of Sciences of the United States of America,1966,55(1):134-141.
[20] ICHISHIMA E.Aspergillopepsin I[M].Handbook of Proteolytic Enzymes.Amsterdam:Elsevier,2013.
[21] KOLATTUKUDY P E,LEE J D,ROGERS L M,et al.Evidence for possible involvement of an elastolytic serine protease in aspergillosis[J].Infection and Immunity,1993,61(6):2 357-2 368.
[22] 赵国忠, 姚云平,曹小红,等.2种米曲霉发酵酱油风味物质比较[J].食品科学,2014,35(24):249-253.
ZHAO G Z,YAO Y P,CAO X H,et al.Comparison of flavor compounds in soy sauces fermented by two Aspergillus oryzae strains[J].Food Science,2014,35(24):249-253.
[23] HAN B Z,ROMBOUTS F M,NOUT M J R.Amino acid profiles of sufu,a Chinese fermented soybean food[J].Journal of Food Composition & Analysis,2004,17(6):689-698.
[24] POTERA C.Making succinate more successful[J].Environmental Health Perspectives,2005,113(12):A832-A835.
[25] PERKINS-VEAZIE P,COLLINS J K.Contributions of nonvolatile phytochemicals to nutrition and flavor[J].Hort Technology,2001,11(4):539-546.
[26] 童佳. 米曲霉发酵高盐稀态酱油过程中挥发性风味物质及蛋白酶表达规律研究[D].无锡:江南大学,2017.
TONG J.Volatile flavor compounds and protease of high-salt liquid-state soy sauce fermentated by Aspergillus oryzae [D].Wuxi:Jiangnan University,2017.
[27] DIEZ-SIMON C,EICHELSHEIM C,MUMM R,et al.Chemical and sensory characteristics of soy sauce:A review[J]. Journal of Agricultural and Food Chemistry,2020,68(42),11 612-11 630.
Outlines

/