固态发酵下酵母自溶的工艺优化
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摘要
固态下酵母自溶可以有效促进菌体内多种活性物质的释放,进而提高酵母类产品的品质。通过优化自溶温度、自溶时间及自溶促进剂锌离子浓度以获得固态发酵下酵母自溶的最佳工艺,对固态发酵物料中游离氨基酸、可溶性蛋白、α-氨基氮含量和A_(260)/A_(280)等指标的分析来确定固态酵母自溶工艺条件,在此基础上以自溶温度40℃、50℃、55℃;作用时间12、18、24 h;锌离子添加浓度2、4、8 mg/kg设置L_9(3~3)正交试验,进一步优化固态酵母自溶的工艺参数。结果表明酵母自溶的最佳工艺条件为:自溶温度55℃、作用时间18 h、锌离子浓度2 mg/kg,此时其可溶性蛋白含量可达9.31 mg/g、游离氨基酸14.36 mg/g、α-氨基氮10.16μg/g、A_(260)/A_(280)为1.73。经工艺优化后,可显著提高酵母自溶产物可溶性蛋白、游离氨基酸和α-氨基氮的含量,从而明显提高了复合菌培养物的品质。
Yeast autolysis under solid-state fermentation can effectively promote the release of various active substances,thereby improving the quality of yeast products. The optimal process for yeast autolysis under solid-state fermentation was obtained by optimizing the autolysis temperature, autolysis time and the zinc ion concentration. We analyzed the indexes of free amino acid, soluble protein and α-amino nitrogen in the fermentation material, as well as A_(260)/A_(280) ratio to determine yeast autolysis process conditions in the solid-state fermentation. On the basis of the obtained data, L_9(3~3) orthogonal test was designed to optimize the solid-state fermentation parameters for yeast autolysis: temperature at 40, 50 and 55℃; time 12, 18 and 24 h; zinc ion concentration 2, 4 and 8 mg/kg. The optimum process conditions for yeast autolysis were: autolysis temperature 55℃, time 18 h, zinc ion concentration 2 mg/kg, and soluble protein content reached 9.31 mg/g, free amino acid 14.36 mg/g,α-amino nitrogen 10.16 μg/g and A_(260)/A_(280) 1.73. After optimization of the process, the soluble protein, free amino acid andα-amino nitrogen contents of the yeast autolysis production can be significantly increased, thereby obviously improving the quality of the composite culture.
Yeast autolysis under solid-state fermentation can effectively promote the release of various active substances,thereby improving the quality of yeast products. The optimal process for yeast autolysis under solid-state fermentation was obtained by optimizing the autolysis temperature, autolysis time and the zinc ion concentration. We analyzed the indexes of free amino acid, soluble protein and α-amino nitrogen in the fermentation material, as well as A_(260)/A_(280) ratio to determine yeast autolysis process conditions in the solid-state fermentation. On the basis of the obtained data, L_9(3~3) orthogonal test was designed to optimize the solid-state fermentation parameters for yeast autolysis: temperature at 40, 50 and 55℃; time 12, 18 and 24 h; zinc ion concentration 2, 4 and 8 mg/kg. The optimum process conditions for yeast autolysis were: autolysis temperature 55℃, time 18 h, zinc ion concentration 2 mg/kg, and soluble protein content reached 9.31 mg/g, free amino acid 14.36 mg/g,α-amino nitrogen 10.16 μg/g and A_(260)/A_(280) 1.73. After optimization of the process, the soluble protein, free amino acid andα-amino nitrogen contents of the yeast autolysis production can be significantly increased, thereby obviously improving the quality of the composite culture.
引文
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[9]Jiang XW,Luo XM,Liu YL.Study on autolysis conditions of spent brewer yeast by changing temperature.Sci Technol Food Ind,2003,24(4):48-50(in Chinese).蒋雪薇,罗晓明,刘永乐.啤酒废酵母变温自溶条件的研究.食品工业科技,2003,24(4):48-50.
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[11]Zhu ZF.Roles of zinc ions in beer brewing and its control.Liquor-Mak Sci Technol,2003,(6):65-66,64(in Chinese).祝忠付.锌离子在啤酒酿造中的作用与控制.酿酒科技,2003,(6):65-66,64.
[12]Zhang JL.Effect of turned material process and different strains of combination on nutricines content of compound bacteria solid state fermentation[D].Hohhot:Inner Mongolia Agricultural University,2015(in Chinese).张锦玲.翻料工艺及不同菌种组合对复合菌固态发酵活性物质含量的影响[D].呼和浩特:内蒙古农业大学,2015.
[13]Guo P,Zhang LM,Zhang Z,et al.Study on the microbial fermentation feed formula and its turning process of dairy cows.China Anim Husband Veter Med,2016,43(10):2598-2607(in Chinese).郭鹏,张立明,张政,等.奶牛微生物发酵饲料组方及其配套翻料工艺的研究.中国畜牧兽医,2016,43(10):2598-2607.
[14]Ibano?lu S.Influence of tempering with ozonated water on the selected properties of wheat flour.J Eng,2001,48(4):345-350.
[15]Gaou I,Dubois M,Pfohl-Leszkowicz A,et al.Safety of Oxygreen?,an ozone treatment on wheat grains.Part 1.A four-week toxicity study in rats by dietary administration of treated wheat.Food Addit Contam,2005,22(11):1113-1119.
[16]Xu WN.Study on the evaluation and mechanism of autolysis in lager brewer’s yeasts[D].Wuxi:Jiangnan University,2014(in Chinese).许维娜.啤酒酵母自溶评价及机理研究[D].无锡:江南大学,2014.
[17]Li HM,Peng SM,Li L,et al.Studies on four conventional methods for protein determination.Chin JBiochem Pharm,2008,29(4):277-278(in Chinese).李海玲,彭书明,李凛,等.4种常用蛋白浓度测定方法的比较.中国生化药物杂志,2008,29(4):277-278.
[18]National Development and Reform Commission of the People’s Republic of China.QB/T1686-2008 Barley malt.Beijing:China Light Industry Press,2008(in Chinese).中华人民共和国国家发展和改革委员会.QB/T1686-2008啤酒麦芽.北京:中国轻工业出版社,2008.
[19]Wang EZ.Method for measuring amino acid concentration and amino acid diagnose reagent kit:CN,CN101082569A.2007-12-05(in Chinese).王尔中.氨基酸浓度的测定方法及氨基酸诊断试剂盒:中国,CN101082569A.2007-12-05.
[20]Sun XJ,Yao XM,Zhang W.A study on condition of yeast autolysis.J Shanghai Agric College,2000,18(1):36-39(in Chinese).孙向军,姚晓敏,张伟.酵母细胞自溶条件的研究.上海农学院学报,2000,18(1):36-39.
[21]Li X,Peng L,Wang Y,et al.The study of yeast autolysis.China Brew,2001,20(5):17-19(in Chinese).李祥,彭莉,王毅,等.酵母自溶研究.中国酿造,2001,20(5):17-19.
[22]Tian XQ,Tu JX,Li HP.Study on autolysis of amino acid solution from waste beer yeast.J Beer Sci Technol,2011,(7):27-29(in Chinese).田小群,涂京霞,李惠萍.啤酒废酵母自溶制备氨基酸液的研究.啤酒科技,2011,(7):27-29.
[23]Tang WX.Study on autolysis conditions of waste brewer’s yeast.Sichuan Food Ferment Technol,2006,42(5):31-33(in Chinese).汤务霞.啤酒废酵母自溶条件的研究.四川食品与发酵,2006,42(5):31-33.
[24]Eide DJ.Zinc transporters and the cellular trafficking of zinc.Biochim Biophys Acta,2006,1763(7):711-722.
[25]de Nicola R,Walker GM.Accumulation and cellular distribution of zinc by brewing yeast.Enzyme Microb Technol,2009,44(4):210-216.
[26]Bromberg SK,Bower PA,Duncombe GR,et al.Requirements for zinc,manganese,calcium,and magnesium in wort.J Am Soc Brew Chem,1997,55(3):123-128.
[27]Li DN.On the environmental factors leading to yeast death and autolysis in beer fermentation process.Sci Technol Innovation Herald,2008,(5):153(in Chinese).李德宁.论啤酒发酵过程中导致酵母死亡及自溶的环境因素.科技创新导报,2008,(5):153.
[28]Wang ZJ,Yu XH,Fang WM.Effects of wort containing abundant ferrum and zinc on metabolic by product of beer yeast.Liquor-Mak Sci Technol,2003,(5):73-76(in Chinese).汪志君,余晓红,方维明.富铁、富锌麦汁对啤酒酵母代谢副产物的影响.酿酒科技,2003,(5):73-76.
[29]Jing XL.Effects of constant and trace ions in brewing water on beer brewing[D].Lanzhou:Gansu Agricultural University,2006(in Chinese).景晓莉.酿造水中的常量离子、微量离子对啤酒酿造的影响[D].兰州:甘肃农业大学,2006.
[30]Jin LQ.Biochemistry.Beijing:People’s Health Publishing House,2007(in Chinese).金丽琴.生物化学.北京:人民卫生出版社,2007.
[2]Premsler T,Zahedi RP,Lewandrowski U,et al.Recent advances in yeast organelle and membrane proteomics.Proteomics,2009,9(20):4731-4743.
[3]Borchani C,Fonteyn F,Jamin G,et al.Enzymatic process for the fractionation of baker’s yeast cell wall(Saccharomyces cerevisiae).Food Chem,2014,163:108-113.
[4]Yang JM,Li H,Du JH.Autolysis conditions of waste brewer’s yeast.China Brew,2012,31(2):95-99(in Chinese).杨建梅,李红,杜金华.啤酒废酵母自溶条件的研究.中国酿造,2012,31(2):95-99.
[5]Zhao ZY,Du JH.The effect of zinc ion addition on yeast autolysis in beer.Liquor Mak,2015,42(6):71-76(in Chinese).赵志云,杜金华.添加锌离子对啤酒酵母自溶的影响.酿酒,2015,42(6):71-76.
[6]Li Y,Bao QB,Kong L,et al.Optimums of food-grade yeast extract processing technology.Food Ind,2011,(5):62-65(in Chinese).李杨,包清彬,孔凌,等.酵母抽提物自溶工艺优化.食品工业,2011,(5):62-65.
[7]Zeng D,Huang JM.Studying of yeast extrac used in dissolved substances from the Pichia pastoris.J Ferment Technol,2011,40(3):9-11(in Chinese).曾栋,黄金明.毕赤酵母自溶物酵母抽提物的研究.发酵科技通讯,2011,40(3):9-11.
[8]Yu Y,Chen Y,Li Q,et al.Optimization of Debaryomyces hansenii autolysis condition through wastewater from monosodium glutamate.Sci Technol Food Ind,2017,38(12):140-145,150(in Chinese).喻轶,陈园,黎琪,等.味精废水培养汉逊德巴利酵母的自溶条件优化.食品工业科技,2017,38(12):140-145,150.
[9]Jiang XW,Luo XM,Liu YL.Study on autolysis conditions of spent brewer yeast by changing temperature.Sci Technol Food Ind,2003,24(4):48-50(in Chinese).蒋雪薇,罗晓明,刘永乐.啤酒废酵母变温自溶条件的研究.食品工业科技,2003,24(4):48-50.
[10]Jiang H,Li HJ,Zhang JJ.Effect of zinc ion on beer brewing.Beer Sci Technol,2014,(7):32-34(in Chinese).蒋虹,李红捷,张京菁.锌离子对啤酒酿造的影响.啤酒科技,2014,(7):32-34.
[11]Zhu ZF.Roles of zinc ions in beer brewing and its control.Liquor-Mak Sci Technol,2003,(6):65-66,64(in Chinese).祝忠付.锌离子在啤酒酿造中的作用与控制.酿酒科技,2003,(6):65-66,64.
[12]Zhang JL.Effect of turned material process and different strains of combination on nutricines content of compound bacteria solid state fermentation[D].Hohhot:Inner Mongolia Agricultural University,2015(in Chinese).张锦玲.翻料工艺及不同菌种组合对复合菌固态发酵活性物质含量的影响[D].呼和浩特:内蒙古农业大学,2015.
[13]Guo P,Zhang LM,Zhang Z,et al.Study on the microbial fermentation feed formula and its turning process of dairy cows.China Anim Husband Veter Med,2016,43(10):2598-2607(in Chinese).郭鹏,张立明,张政,等.奶牛微生物发酵饲料组方及其配套翻料工艺的研究.中国畜牧兽医,2016,43(10):2598-2607.
[14]Ibano?lu S.Influence of tempering with ozonated water on the selected properties of wheat flour.J Eng,2001,48(4):345-350.
[15]Gaou I,Dubois M,Pfohl-Leszkowicz A,et al.Safety of Oxygreen?,an ozone treatment on wheat grains.Part 1.A four-week toxicity study in rats by dietary administration of treated wheat.Food Addit Contam,2005,22(11):1113-1119.
[16]Xu WN.Study on the evaluation and mechanism of autolysis in lager brewer’s yeasts[D].Wuxi:Jiangnan University,2014(in Chinese).许维娜.啤酒酵母自溶评价及机理研究[D].无锡:江南大学,2014.
[17]Li HM,Peng SM,Li L,et al.Studies on four conventional methods for protein determination.Chin JBiochem Pharm,2008,29(4):277-278(in Chinese).李海玲,彭书明,李凛,等.4种常用蛋白浓度测定方法的比较.中国生化药物杂志,2008,29(4):277-278.
[18]National Development and Reform Commission of the People’s Republic of China.QB/T1686-2008 Barley malt.Beijing:China Light Industry Press,2008(in Chinese).中华人民共和国国家发展和改革委员会.QB/T1686-2008啤酒麦芽.北京:中国轻工业出版社,2008.
[19]Wang EZ.Method for measuring amino acid concentration and amino acid diagnose reagent kit:CN,CN101082569A.2007-12-05(in Chinese).王尔中.氨基酸浓度的测定方法及氨基酸诊断试剂盒:中国,CN101082569A.2007-12-05.
[20]Sun XJ,Yao XM,Zhang W.A study on condition of yeast autolysis.J Shanghai Agric College,2000,18(1):36-39(in Chinese).孙向军,姚晓敏,张伟.酵母细胞自溶条件的研究.上海农学院学报,2000,18(1):36-39.
[21]Li X,Peng L,Wang Y,et al.The study of yeast autolysis.China Brew,2001,20(5):17-19(in Chinese).李祥,彭莉,王毅,等.酵母自溶研究.中国酿造,2001,20(5):17-19.
[22]Tian XQ,Tu JX,Li HP.Study on autolysis of amino acid solution from waste beer yeast.J Beer Sci Technol,2011,(7):27-29(in Chinese).田小群,涂京霞,李惠萍.啤酒废酵母自溶制备氨基酸液的研究.啤酒科技,2011,(7):27-29.
[23]Tang WX.Study on autolysis conditions of waste brewer’s yeast.Sichuan Food Ferment Technol,2006,42(5):31-33(in Chinese).汤务霞.啤酒废酵母自溶条件的研究.四川食品与发酵,2006,42(5):31-33.
[24]Eide DJ.Zinc transporters and the cellular trafficking of zinc.Biochim Biophys Acta,2006,1763(7):711-722.
[25]de Nicola R,Walker GM.Accumulation and cellular distribution of zinc by brewing yeast.Enzyme Microb Technol,2009,44(4):210-216.
[26]Bromberg SK,Bower PA,Duncombe GR,et al.Requirements for zinc,manganese,calcium,and magnesium in wort.J Am Soc Brew Chem,1997,55(3):123-128.
[27]Li DN.On the environmental factors leading to yeast death and autolysis in beer fermentation process.Sci Technol Innovation Herald,2008,(5):153(in Chinese).李德宁.论啤酒发酵过程中导致酵母死亡及自溶的环境因素.科技创新导报,2008,(5):153.
[28]Wang ZJ,Yu XH,Fang WM.Effects of wort containing abundant ferrum and zinc on metabolic by product of beer yeast.Liquor-Mak Sci Technol,2003,(5):73-76(in Chinese).汪志君,余晓红,方维明.富铁、富锌麦汁对啤酒酵母代谢副产物的影响.酿酒科技,2003,(5):73-76.
[29]Jing XL.Effects of constant and trace ions in brewing water on beer brewing[D].Lanzhou:Gansu Agricultural University,2006(in Chinese).景晓莉.酿造水中的常量离子、微量离子对啤酒酿造的影响[D].兰州:甘肃农业大学,2006.
[30]Jin LQ.Biochemistry.Beijing:People’s Health Publishing House,2007(in Chinese).金丽琴.生物化学.北京:人民卫生出版社,2007.
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