脂肪酶处理白酒丢糟提升复糟酒的品质
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摘要
本项目以浓香型白酒丢糟为研究对象,研究巴氏灭菌、反应温度、食用酒精添加量、南极假丝酵母脂肪酶B(Candida antarctica lipase B,CALB)及反应时间等工艺条件对复糟酒酯化效果的影响。实验结果表明,经巴氏灭菌处理后,在食用酒精(95%,V/V)用量为20 mL/100 g丢糟、CALB酶添加量1.5‰(W/W)、20℃下反应3 d,增酯增香的效果最好,乙酸乙酯、己酸乙酯、乳酸乙酯及总酯含量依次为3.17 g/L、0.59 g/L、0.76 g/L、4.53 g/L。在此基础上进行1 kg酒糟处理量小试实验,得乙酸乙酯、己酸乙酯、乳酸乙酯及总酯含量依次为3.44 g/L、0.85 g/L、0.70 g/L和4.98 g/L,其中浓香型白酒的主要呈香物质乙酸乙酯和己酸乙酯分别较无CALB酶处理实验组提高了16.38倍和16.67倍,且放大前后各酯含量基本保持稳定。本研究为进一步放大实验提供了依据,对脂肪酶催化白酒丢糟资源化再生利用具有重要意义。
In this study, distiller's grains of Luzhou-flavor liquor were used as the research object, and the effects of processing conditions such as pasteurization, reaction temperature, edible alcohol addition, Candida antarctica lipase B(CALB) and reaction time on alcohol esterification were studied. The experimental results showed that the greatest enhancement on ester content and flavor occurred after pasteurization and reactions under the following conditions: Dose of edible alcohol(95%, V/V), 20 mL/100 g; amount of CALB enzyme, 1.5 ‰(W/W), reaction temperature and time, 20 ℃ and 3 days. Under which, the contents of ethyl acetate, ethyl hexanoate, ethyl lactate and total ester were 3.17 g/L, 0.59 g/L, 0.76 g/L and 4.53 g/L in the product, respectively. Accordingly, further experiments using 1 kg of distiller's grains were carried out, and the contents of ethyl acetate, ethyl hexanoate, ethyl lactate and total ester were 3.44 g/L, 0.85 g/L, 0.70 g/L and 4.98 g/L,respectively. The amounts of the main aroma substances of Luzhou-flavor liquor, ethyl acetate and ethyl hexanoate, in the CALB-treated sample were increased by 16.38 times and 16.67 times, respectively, compared to those without CALB enzyme treatment, and the contents of individual esters remained constant after amplification. This study provides a basis for further amplification experiments, and is of great significance to lipase-catalyzed recycling of distiller's grains.
In this study, distiller's grains of Luzhou-flavor liquor were used as the research object, and the effects of processing conditions such as pasteurization, reaction temperature, edible alcohol addition, Candida antarctica lipase B(CALB) and reaction time on alcohol esterification were studied. The experimental results showed that the greatest enhancement on ester content and flavor occurred after pasteurization and reactions under the following conditions: Dose of edible alcohol(95%, V/V), 20 mL/100 g; amount of CALB enzyme, 1.5 ‰(W/W), reaction temperature and time, 20 ℃ and 3 days. Under which, the contents of ethyl acetate, ethyl hexanoate, ethyl lactate and total ester were 3.17 g/L, 0.59 g/L, 0.76 g/L and 4.53 g/L in the product, respectively. Accordingly, further experiments using 1 kg of distiller's grains were carried out, and the contents of ethyl acetate, ethyl hexanoate, ethyl lactate and total ester were 3.44 g/L, 0.85 g/L, 0.70 g/L and 4.98 g/L,respectively. The amounts of the main aroma substances of Luzhou-flavor liquor, ethyl acetate and ethyl hexanoate, in the CALB-treated sample were increased by 16.38 times and 16.67 times, respectively, compared to those without CALB enzyme treatment, and the contents of individual esters remained constant after amplification. This study provides a basis for further amplification experiments, and is of great significance to lipase-catalyzed recycling of distiller's grains.
引文
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[12]马文鹏,裴芳霞,任海伟,等.双酶复合水解酒糟制备可发酵糖的工艺研究[J].酿酒科技,2016,4:89-92MA Wen-peng,PEI Fang-xia,REN Hai-wei,et al.The preparation of fermentable sugar through the hydrolysis of distillers grains by two enzymes[J].Liquor-Making Science&Technology,2016,4:89-92
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[14]任海伟,徐娜,李金平,等.化学预处理提高酒糟生物质酶解糖化效果[J].农业工程学报,2014,30(16):239-246REN Hai-wei,XU Na,LI Jin-ping,et al.Chemical pretreatment improving effect of enzymatic saccharification of distillers grains biomass[J].Transactions of the Chinese Society of Agricultural Engineering,2014,30(16):239-246
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[16]王月梅,赵迎路.清香型白酒发酵中的酯化研究[J].酿酒科技,2003,1:47-49WANG Yue-mei,ZHAO Ying-lu.Study on the esterification in the fermentation of fen-flavor liquors[J].Liquor-making Science&Technology,2003,1:47-49
[17]Larios A,García H S,Oliart R M,et al.Synthesis of flavor and fragrance esters using Candida antarctica lipase[J].Applied Microbiology&Biotechnology,2004,65(4):373-376
[18]赵东,彭志云,牛广杰,等.强化发酵丢糟再生产白酒的研究[J].中国酿造,2011,30(2):147-149ZHAO Dong,PENG Zhi-yun,NIU Guang-jie,et al.Recycle and utilization of distiller's grains for Chinese liquor production by fortified fermentation[J].China Brewing,2011,30(2):147-149
[19]任道群,唐玉明,姚万春,等.酯化酶动力学研究[J].酿酒科技,2006,6:39-40REN Dao-qun,TANG Yu-ming,YAO Wan-chun,et al.Research on the kinetics of esterifying enzyme[J].Liquor-Making Science&Technology,2006,6:39-40
[20]崔如生,徐岩.应用R92华根霉生产调味酒提高新型白酒质量[J].酿酒,2007,34(1):27-31CUI Ru-sheng,YAN XU.Producing flavoring liquid by using rhizopus Chinese R92 to improve new type liquor quality[J].Liquor Making,2007,34(1):27-31
[21]窦晓,杨建刚,马莹莹,等.华根霉在酿酒中的研究进展[J].食品与发酵工业,2015,41(4):246-250DOU Xiao,YANG Jian-gang,MA Ying-ying,et al.The research progress about Rhizopus chinonsis in liquor-making[J].Food&Fermentation Industries,2015,41(4):246-250
[22]颜兴和,王栋,徐岩.华根霉脂肪酶有机相合成酶活的研究[J].工业微生物,2005,35(2):24-28YAN Xing-He,WANG Dong,XU Yan.Synthetic activity of Rhizopus chinensis lipase in organic media[J].Industrial Microbiology,2005,35(2):24-28
[23]Xu Y,Wang D,Mu X Q,et al.Biosynthesis of ethyl esters of short-chain fatty acids using whole-cell lipase from Rhizopus chinensis,CCTCC M201021 in non-aqueous phase[J].Journal of Molecular Catalysis B Enzymatic,2002,18(1):29-37
[24]Sun S Y,Xu Y.Solid-state fermentation for'whole-cell synthetic lipase'production from Rhizopus chinensis and identification of the functional enzyme[J].Process Biochemistry,2008,43(2):219-224
[25]潘志友,韩双艳,林影,等.南极假丝酵母脂肪酶B的酿酒酵母表面展示及其催化已酸乙酯的合成[J].生物工程学报,2008,24(4):673-678PAN Zhi-you,HAN Shuang-yan,LIN Ying,et al.Expression of Candida antarctica lipase B on yeast surface and synthesis of ethyl hexanoate catalyzed by CALB[J].Chinese Journal of Biotechnology,2008,24(4):673-678
[26]赵天涛,高静,李伟杰.南极假丝酵母脂肪酶B的催化机理及应用前景[J].分子催化,2005,9(2):155-160ZHAO Tian-tao,GAO Jing,LI Wei-jie,et al.The catalytic mechanism and application of Candida antarctic lipase B[J].Journal of Molecular Catalysis,2005,19(2):155-160
[27]张娜,金子,林影,等.酵母表面展示脂肪酶合成己二酸二异辛酯[J].生物工程学报,2013,29(7):1027-1031ZHANG Na,JIN Zi,LIN Ying,et al.Synthesis of diisooctyl adipate catalyzed by lipase-displaying Pichia pastoris whole-cell biocatalysts[J].Chinese Journal of Biotechnology,2013,29(7):1027-1031
[28]金子,林影,黄登峰,等.展示南极假丝酵母脂肪酶B的毕赤酵母全细胞催化合成短链芳香酯[J].生物工程学报,2009,25(12):1927-1932JIN Zi,LIN Ying,HUANG Deng-feng,et al.Synthesis of flavor esters catalyzed by CALB-displaying Pichia pastoris whole-cells in non-aqueous phase[J].Chinese Journal of Biotechnology,2009,25(12):1927-1932
[29]JIN Zi,Janvier Ntwali,LIN Ying,et al.Geranyl Butyrate Production by Candida antarctica,lipase B-Displaying Pichia pastoris[M].Advances in Applied Biotechnology.Springer Berlin Heidelberg,2015
[30]孙海龙.脂肪酶处理清香型白酒丢糟生成香气物质的研究[D].石家庄:河北科技大学,2013SUN Hai-long.Research of the lipase used on the esterification of organic acid in Fng Liqutor’s Lees.[D].Shijiazhuang:Hebei University of Science and Technology,2013
[2]李芳香,张稳,郁建平,等.茅台酱香型酒糟基本成分的测定与分析[J].贵州农业科学,2016,44(9):114-116LI Fang-xiang,ZHANG Wen,YU Jian-ping,et al.Determination and analysis of basic components of vinasse with moutai-flavor type[J].Guizhou Agricultural Sciences,2016,44(9):114-116
[3]方晓璞,张文学,张其圣,等.丢糟酿酒复合发酵剂的应用开发研究[J].中国酿造,2007,2007(4):55-57FANG Xiao-pu,ZHANG Wen-xue,ZHANG Qi-sheng,et al.Application and development of compound starters for liquor fermentation from waste lees[J].China Brewing,2007,2007(4):55-57
[4]张丽华,王小媛,李昌文,等.酒糟再利用的研究进展[J].食品与发酵工业,2017,43(11):250-256ZHANG Li-hua,WANG Xiao-yuan,LI Chang-wen,et al.Recent advances in comprehensive utilization of grain stillage[J].Food and Fermentation Industries,2007,43(11):250-256
[5]李政一,周定,侯文华.酒糟资源化研究[J].环境科学学报,2000,s1:145-149LI Zheng-yi,ZHOU Ding,HOU Wen-hua.Studies on the resource recovery technique of distillers grain[J].Acta Scientiae Circumstantiae,2000,s1:145-149
[6]袁颉,邱树毅,彭正东,等.固态白酒生产酒糟的资源化利用研究进展[J].酿酒科技,2012,5:88-91YUAN Jie,QIU Shu-yi,PENG Zheng-dong,et al.Research advance in the reclamation of distiller’s grains in liquor production by solid fermentation[J].Liquor-Making Science&Technology,2012,5:88-91
[7]高路.酒糟的综合利用[J].酿酒科技,2004,5:101-102GAO Lu.Comprehensive utilization of distiller’s grains[J].Liquor-making Science&Technology,2004,5:101-102
[8]张敏.酒糟制甘油的生产工艺及研究进展[J].湖南科技学院学报,2001,22(3):47-49ZHANG Min.Production of glycerol with distillers,grains and research progress[J].Journal of Hunan University of Science and Engineering,2001,22(3):47-49
[9]任羽,王松,王涛.酒糟栽培食用菌研究现状[J].中国酿造,2017,36(3):5-9REN Yu,WANG Song,WANG Tao.Research status of edible fungi cultivated by distillers'grains[J].China Brewing,2017,36(3):5-9
[10]陈佳兴,邱树毅,王雪郦.醇碱法提取酱香型白酒酒糟中蛋白质的工艺研究[J].食品工业,2018,39(2):37-40CHEN Jia-xing,QIU Shu-yi,WANG Xue-li.Study on the extraction of protein from wine lees of maotai-flavor liquor by alcohol-alkali method[J].Food Industry,2018,39(2):37-40
[11]范文权,高洪涛.鲜酒糟加工食醋工艺研究[J].酿酒科技,2000,5:89-89FAN Wen-quan,GAO Hong-tao.Technological research on the production of vinegar with fresh distiller′s grains[J].Liquor-Making Science&Technology,2000,5:89-89
[12]马文鹏,裴芳霞,任海伟,等.双酶复合水解酒糟制备可发酵糖的工艺研究[J].酿酒科技,2016,4:89-92MA Wen-peng,PEI Fang-xia,REN Hai-wei,et al.The preparation of fermentable sugar through the hydrolysis of distillers grains by two enzymes[J].Liquor-Making Science&Technology,2016,4:89-92
[13]刘高梅,陈海秀,任海伟.酒糟酶解糖化条件的研究[J].食品工业科技,2012,33(15):138-140LIU Gao-mei,CHEN Hai-xiu,REN Hai-wei.Distiller’s grains pretreatment enzymatic saccharification reducing sugar[J].Science and Technology of Food Industry,2012,33(15):138-140
[14]任海伟,徐娜,李金平,等.化学预处理提高酒糟生物质酶解糖化效果[J].农业工程学报,2014,30(16):239-246REN Hai-wei,XU Na,LI Jin-ping,et al.Chemical pretreatment improving effect of enzymatic saccharification of distillers grains biomass[J].Transactions of the Chinese Society of Agricultural Engineering,2014,30(16):239-246
[15]Bi D X,Chu D Q,Zhu P,et al.Utilization of dry distiller's grain and solubles as nutrient supplement in the simultaneous saccharification and ethanol fermentation at high solids loading of corn stover[J].Biotechnology Letters,2011,33(2):273-276
[16]王月梅,赵迎路.清香型白酒发酵中的酯化研究[J].酿酒科技,2003,1:47-49WANG Yue-mei,ZHAO Ying-lu.Study on the esterification in the fermentation of fen-flavor liquors[J].Liquor-making Science&Technology,2003,1:47-49
[17]Larios A,García H S,Oliart R M,et al.Synthesis of flavor and fragrance esters using Candida antarctica lipase[J].Applied Microbiology&Biotechnology,2004,65(4):373-376
[18]赵东,彭志云,牛广杰,等.强化发酵丢糟再生产白酒的研究[J].中国酿造,2011,30(2):147-149ZHAO Dong,PENG Zhi-yun,NIU Guang-jie,et al.Recycle and utilization of distiller's grains for Chinese liquor production by fortified fermentation[J].China Brewing,2011,30(2):147-149
[19]任道群,唐玉明,姚万春,等.酯化酶动力学研究[J].酿酒科技,2006,6:39-40REN Dao-qun,TANG Yu-ming,YAO Wan-chun,et al.Research on the kinetics of esterifying enzyme[J].Liquor-Making Science&Technology,2006,6:39-40
[20]崔如生,徐岩.应用R92华根霉生产调味酒提高新型白酒质量[J].酿酒,2007,34(1):27-31CUI Ru-sheng,YAN XU.Producing flavoring liquid by using rhizopus Chinese R92 to improve new type liquor quality[J].Liquor Making,2007,34(1):27-31
[21]窦晓,杨建刚,马莹莹,等.华根霉在酿酒中的研究进展[J].食品与发酵工业,2015,41(4):246-250DOU Xiao,YANG Jian-gang,MA Ying-ying,et al.The research progress about Rhizopus chinonsis in liquor-making[J].Food&Fermentation Industries,2015,41(4):246-250
[22]颜兴和,王栋,徐岩.华根霉脂肪酶有机相合成酶活的研究[J].工业微生物,2005,35(2):24-28YAN Xing-He,WANG Dong,XU Yan.Synthetic activity of Rhizopus chinensis lipase in organic media[J].Industrial Microbiology,2005,35(2):24-28
[23]Xu Y,Wang D,Mu X Q,et al.Biosynthesis of ethyl esters of short-chain fatty acids using whole-cell lipase from Rhizopus chinensis,CCTCC M201021 in non-aqueous phase[J].Journal of Molecular Catalysis B Enzymatic,2002,18(1):29-37
[24]Sun S Y,Xu Y.Solid-state fermentation for'whole-cell synthetic lipase'production from Rhizopus chinensis and identification of the functional enzyme[J].Process Biochemistry,2008,43(2):219-224
[25]潘志友,韩双艳,林影,等.南极假丝酵母脂肪酶B的酿酒酵母表面展示及其催化已酸乙酯的合成[J].生物工程学报,2008,24(4):673-678PAN Zhi-you,HAN Shuang-yan,LIN Ying,et al.Expression of Candida antarctica lipase B on yeast surface and synthesis of ethyl hexanoate catalyzed by CALB[J].Chinese Journal of Biotechnology,2008,24(4):673-678
[26]赵天涛,高静,李伟杰.南极假丝酵母脂肪酶B的催化机理及应用前景[J].分子催化,2005,9(2):155-160ZHAO Tian-tao,GAO Jing,LI Wei-jie,et al.The catalytic mechanism and application of Candida antarctic lipase B[J].Journal of Molecular Catalysis,2005,19(2):155-160
[27]张娜,金子,林影,等.酵母表面展示脂肪酶合成己二酸二异辛酯[J].生物工程学报,2013,29(7):1027-1031ZHANG Na,JIN Zi,LIN Ying,et al.Synthesis of diisooctyl adipate catalyzed by lipase-displaying Pichia pastoris whole-cell biocatalysts[J].Chinese Journal of Biotechnology,2013,29(7):1027-1031
[28]金子,林影,黄登峰,等.展示南极假丝酵母脂肪酶B的毕赤酵母全细胞催化合成短链芳香酯[J].生物工程学报,2009,25(12):1927-1932JIN Zi,LIN Ying,HUANG Deng-feng,et al.Synthesis of flavor esters catalyzed by CALB-displaying Pichia pastoris whole-cells in non-aqueous phase[J].Chinese Journal of Biotechnology,2009,25(12):1927-1932
[29]JIN Zi,Janvier Ntwali,LIN Ying,et al.Geranyl Butyrate Production by Candida antarctica,lipase B-Displaying Pichia pastoris[M].Advances in Applied Biotechnology.Springer Berlin Heidelberg,2015
[30]孙海龙.脂肪酶处理清香型白酒丢糟生成香气物质的研究[D].石家庄:河北科技大学,2013SUN Hai-long.Research of the lipase used on the esterification of organic acid in Fng Liqutor’s Lees.[D].Shijiazhuang:Hebei University of Science and Technology,2013