解淀粉芽孢杆菌GSBa-1凝乳酶在马苏里拉干酪中的应用
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摘要
对分离自酒曲的1株解淀粉芽孢杆菌GSBa-1发酵所产凝乳酶进行研究,该酶凝乳活力高而蛋白水解活力低,纯酶凝乳活力可达1.46×106 SU/g;使用该凝乳酶和商品凝乳酶制作马苏里拉干酪,并对干酪理化成分、成熟过程中pH值和微生物指标及干酪成熟前后质构特性、游离脂肪酸、可溶性蛋白、风味和干酪性能等指标进行对比分析。结果显示,理化成分上菌株凝乳酶与商品凝乳酶制作的干酪相接近(P<0.05)。干酪在成熟过程中,发酵剂存活数先增加后减少,但其差异不大;菌株凝乳酶制作的干酪pH 4.6可溶性蛋白含量较多,干酪的游离氨基酸总量(76 mg/100 g)也高于商品凝乳酶制作的干酪游离氨基酸总量(55.3 mg/100 g);菌株凝乳酶制作的干酪质构特性优于商品凝乳酶制作的干酪;电镜结果显示,菌株凝乳酶制作的干酪内部网状结构更充实;菌株凝乳酶具有稍强的蛋白水解活力,导致其制作的干酪风味物质种类多于商品凝乳酶制作的干酪,风味物质更加丰富。干酪样品的保形性和拉丝性实验测定结果显示,2种凝乳酶制作的干酪性能差异不大(P>0.05);对2种凝乳酶制作的干酪进行感官评定,其总评分相接近。以上结果表明,解淀粉芽孢杆菌GSBa-1凝乳酶在一定程度上可代替小牛凝乳酶应用于马苏里拉干酪的生产。
The rennet produced by Bacillus amyloliquefaciens GSBa-1, isolated from Jiuqu, a traditional Chinese liquor fermentation starter, was found to possess high milk-clotting activity(up to 1.46 × 106 SU/g) and low proteolytic activity.Mozzarella cheeses were prepared using the purified enzyme, and a commercial rennet was used as a control. During cheese ripening, the physicochemical properties, pH, microbial parameters, texture properties, free fatty acid composition, soluble protein content, flavor properties and other properties were studied. Results showed no significant difference(P < 0.05) in chemical composition was seen between the two cheeses. During cheese ripening, the viable count of Lactococcus lactis increased first and then decreased; however, the difference was not significant. The pH 4.6 soluble protein content and total free amino acid content(76 versus 55.3 mg/100 g) of cheese made with the microbial rennet were higher than those of the control cheese. The cheese prepared using the microbial rennet also had better textural properties than the control.Morphological observation of cheese using scanning electron microscopy showed that the internal network structure of the cheese prepared using the microbial rennet was more compact. Moreover, it contained more volatile flavor compounds due to the relatively strong proteolytic activity of the rennet. There were no significant differences in conformality,stringiness or sensory evaluation between the two cheeses(P > 0.05). All these results showed that the rennet produced by B. amyloliquefaciens GSBa-1 is a promising alternative to calf rennet in processing of Mozzarella cheese.
The rennet produced by Bacillus amyloliquefaciens GSBa-1, isolated from Jiuqu, a traditional Chinese liquor fermentation starter, was found to possess high milk-clotting activity(up to 1.46 × 106 SU/g) and low proteolytic activity.Mozzarella cheeses were prepared using the purified enzyme, and a commercial rennet was used as a control. During cheese ripening, the physicochemical properties, pH, microbial parameters, texture properties, free fatty acid composition, soluble protein content, flavor properties and other properties were studied. Results showed no significant difference(P < 0.05) in chemical composition was seen between the two cheeses. During cheese ripening, the viable count of Lactococcus lactis increased first and then decreased; however, the difference was not significant. The pH 4.6 soluble protein content and total free amino acid content(76 versus 55.3 mg/100 g) of cheese made with the microbial rennet were higher than those of the control cheese. The cheese prepared using the microbial rennet also had better textural properties than the control.Morphological observation of cheese using scanning electron microscopy showed that the internal network structure of the cheese prepared using the microbial rennet was more compact. Moreover, it contained more volatile flavor compounds due to the relatively strong proteolytic activity of the rennet. There were no significant differences in conformality,stringiness or sensory evaluation between the two cheeses(P > 0.05). All these results showed that the rennet produced by B. amyloliquefaciens GSBa-1 is a promising alternative to calf rennet in processing of Mozzarella cheese.
引文
[1]李亮,吴丹,宿玲恰,等.重组毕赤酵母生产凝乳酶发酵优化[J].食品与生物技术学报,2016,35(5):457-464.DOI:10.3969/j.issn.1673-1689.2016.05.002.
[2]杭峰,洪青,王钦博,等.凝乳酶的研究进展[J].食品科学,2016,37(3):273-279.DOI:10.7506/spkx1002-6630-201603047.
[3]普燕,张富春.干酪用牛凝乳酶替代品的研究进展[J].食品与发酵工业,2015,41(5):227-234.DOI:10.13995/j.cnki.11-1802/ts.201505041.
[4]潘道东,韩玲玲.根霉凝乳酶的分离纯化及其酶学特性研究[J].中国食品学报,2011,11(2):53-59.DOI:10.3969/j.issn.1009-7848.2011.02.008.
[5]宋曦,甘伯中,贺晓玲,等.天祝放牧牦牛生活环境土壤中一株产凝乳酶细菌的分离与鉴定[J].食品科学,2009,30(11):158-162.DOI:10.3321/j.issn:1002-6630.2009.11.035.
[6]丁明亮,欧阳安然,王望斐,等.枯草芽孢杆菌产凝乳酶发酵条件的优化[J].食品科学,2011,32(3):156-160.
[7]高维东,甘伯中,丁福军,等.微生物凝乳酶的研究进展[J].中国乳品工业,2009,37(5):34-47.DOI:10.3969/j.issn.1001-2230.2009.05.010.
[8]李学朋,关明玲,赵保堂,等.米黑毛霉酶凝干酪素生产工艺参数的优化研究[J].食品工业科技,2016,37(3):191-195.DOI:10.13386/j.issn1002-0306.2016.03.032.
[9]ZHAO X,WANG J,ZHENG Z,et al.Production of a milk-clotting enzyme by glutinous rice fermentation and partial characterization of the enzyme[J].Journal of Food Biochemistry,2015,39(1):70-79.DOI:10.1111/jfbc.12108.
[10]DA SILVA R R,SOUTO T B,DE OLIVEIRA T B,et al.Evaluation of the catalytic specificity,biochemical properties,and milk clotting abilities of an aspartic peptidase from Rhizomucor miehei[J].Journal of Industrial Microbiology&Biotechnology,2016,43(8):1059-1069.DOI:10.1007/s10295-016-1780-4.
[11]李建涛,陈历俊,姜铁民.响应面法优化解淀粉芽孢杆菌发酵产凝乳酶的工艺条件[J].中国乳品工业,2012,40(6):26-30.DOI:10.3969/j.issn.1001-2230.2012.06.007.
[12]腾军伟,赵笑,杨亚威,等.酒曲中产凝乳酶微生物菌株的分离筛选及鉴定[J].食品科学,2017,38(16):23-28.DOI:10.7506/spkx1002-6630-201716004.
[13]腾军伟,杨贞耐.解淀粉芽孢杆菌GSBa-1产凝乳酶培养基组成的优化[J].食品科学技术学报,2016,34(4):31-38.DOI:10.3969/j.issn.2095-6002.2016.04.006.
[14]腾军伟,余志坚,陈超,等.影响解淀粉芽孢杆菌GSBa-1产凝乳酶的因素及发酵条件优化研究[J].中国食品添加剂,2016(8):53-62.DOI:10.3969/j.issn.1006-2513.2016.08.002.
[15]腾军伟,郑喆,梅雪洋,等.解淀粉芽孢杆菌GSBa-1凝乳酶的生产及其酶学性质研究[J].食品与发酵工业,2016,42(12):83-89.DOI:10.13995/j.cnki.11-1802/ts.201612015.
[16]JOHNSON M,LUCEY J.Major technological advances and trends in cheese[J].Journal of Dairy Science,2006,89(4):1174-1178.DOI:10.3168/jds.S0022-0302(06)72186-5.
[17]杨亚威.产胞外多糖乳酸菌的益生特性及其对成熟干酪品质的影响[D].北京:北京工商大学,2016.
[18]赵笑.酒曲中微生物产凝乳酶的特性及其应用研究[D].北京:北京工商大学,2015.
[19]YUN J J,KIELY L J,BARBANO D M,et al.Mozzarella cheese:impact of cooking temperature on chemical composition,proteolysis and functional properties 1[J].Journal of Dairy Science,1993,76(12):3664-3673.DOI:10.3168/jds.S0022-0302(93)77708-5.
[20]FORTIN M H,CHAMPAGNE C P,ST-GELAIS D,et al.Effect of time of inoculation,starter addition,oxygen level and salting on the viability of probiotic cultures during Cheddar cheese production[J].International Dairy Journal,2011,21(2):75-82.DOI:10.1016/j.idairyj.2010.09.007.
[21]DIEZHANDINO I,FERNANDEZ D,GONZALEZ L,et al.Microbiological,physico-chemical and proteolytic changes in a Spanish blue cheese during ripening[J].Food Chemistry,2015,168:134-141.DOI:10.1016/j.foodchem.2014.07.039.
[22]BORSTING M W,OVIST K B,RASMUSSEN M,et al.Impact of selected coagulants and starters on primary proteolysis and amino acid release related to bitterness and structure of reduced-fat Cheddar cheese[J].Dairy Science&Technology,2012,92(5):593-612.
[23]O’MAHONY J A,LUCEY J A,MCSWEENEY P L H.Chymosinmediated proteolysis,calcium solubilization,and texture development during the ripening of Cheddar cheese[J].Journal of Dairy Science,2005,88(9):3101-3114.DOI:10.3168/jds.S0022-0302(05)72992-1.
[24]SOODAM K,ONG L,POWELL I B,et al.Effect of calcium chloride addition and draining pH on the microstructure and texture of full fat Cheddar cheese during ripening[J].Food Chemistry,2015,181:111-118.DOI:10.1016/j.foodchem.2015.01.135.
[25]ONG L,DAGASTINE R R,KENTISH S E,et al.The effect of pH at renneting on the microstructure,composition and texture of Cheddar cheese[J].Food Research International,2012,48(1):119-130.DOI:10.1016/j.foodres.2012.02.020.
[26]艾对,张富新,李延华,等.顶空固态微萃取-气质联用法分析羊乳和牛乳中挥发性化合物[J].食品与发酵工业,2014,40(10):189-193.
[27]WOLF I V,PEROTTI M C,ZALAZAR C A.Composition and volatile profiles of commercial Argentinean blue cheeses[J].Journal of the Science of Food and Agriculture,2011,91(2):385-393.DOI:10.1002/jsfa.4198.
[28]李良.乳酸乳球菌发酵稀奶油过程中甲基酮的合成代谢调控[D].哈尔滨:哈尔滨工业大学,2015.
[2]杭峰,洪青,王钦博,等.凝乳酶的研究进展[J].食品科学,2016,37(3):273-279.DOI:10.7506/spkx1002-6630-201603047.
[3]普燕,张富春.干酪用牛凝乳酶替代品的研究进展[J].食品与发酵工业,2015,41(5):227-234.DOI:10.13995/j.cnki.11-1802/ts.201505041.
[4]潘道东,韩玲玲.根霉凝乳酶的分离纯化及其酶学特性研究[J].中国食品学报,2011,11(2):53-59.DOI:10.3969/j.issn.1009-7848.2011.02.008.
[5]宋曦,甘伯中,贺晓玲,等.天祝放牧牦牛生活环境土壤中一株产凝乳酶细菌的分离与鉴定[J].食品科学,2009,30(11):158-162.DOI:10.3321/j.issn:1002-6630.2009.11.035.
[6]丁明亮,欧阳安然,王望斐,等.枯草芽孢杆菌产凝乳酶发酵条件的优化[J].食品科学,2011,32(3):156-160.
[7]高维东,甘伯中,丁福军,等.微生物凝乳酶的研究进展[J].中国乳品工业,2009,37(5):34-47.DOI:10.3969/j.issn.1001-2230.2009.05.010.
[8]李学朋,关明玲,赵保堂,等.米黑毛霉酶凝干酪素生产工艺参数的优化研究[J].食品工业科技,2016,37(3):191-195.DOI:10.13386/j.issn1002-0306.2016.03.032.
[9]ZHAO X,WANG J,ZHENG Z,et al.Production of a milk-clotting enzyme by glutinous rice fermentation and partial characterization of the enzyme[J].Journal of Food Biochemistry,2015,39(1):70-79.DOI:10.1111/jfbc.12108.
[10]DA SILVA R R,SOUTO T B,DE OLIVEIRA T B,et al.Evaluation of the catalytic specificity,biochemical properties,and milk clotting abilities of an aspartic peptidase from Rhizomucor miehei[J].Journal of Industrial Microbiology&Biotechnology,2016,43(8):1059-1069.DOI:10.1007/s10295-016-1780-4.
[11]李建涛,陈历俊,姜铁民.响应面法优化解淀粉芽孢杆菌发酵产凝乳酶的工艺条件[J].中国乳品工业,2012,40(6):26-30.DOI:10.3969/j.issn.1001-2230.2012.06.007.
[12]腾军伟,赵笑,杨亚威,等.酒曲中产凝乳酶微生物菌株的分离筛选及鉴定[J].食品科学,2017,38(16):23-28.DOI:10.7506/spkx1002-6630-201716004.
[13]腾军伟,杨贞耐.解淀粉芽孢杆菌GSBa-1产凝乳酶培养基组成的优化[J].食品科学技术学报,2016,34(4):31-38.DOI:10.3969/j.issn.2095-6002.2016.04.006.
[14]腾军伟,余志坚,陈超,等.影响解淀粉芽孢杆菌GSBa-1产凝乳酶的因素及发酵条件优化研究[J].中国食品添加剂,2016(8):53-62.DOI:10.3969/j.issn.1006-2513.2016.08.002.
[15]腾军伟,郑喆,梅雪洋,等.解淀粉芽孢杆菌GSBa-1凝乳酶的生产及其酶学性质研究[J].食品与发酵工业,2016,42(12):83-89.DOI:10.13995/j.cnki.11-1802/ts.201612015.
[16]JOHNSON M,LUCEY J.Major technological advances and trends in cheese[J].Journal of Dairy Science,2006,89(4):1174-1178.DOI:10.3168/jds.S0022-0302(06)72186-5.
[17]杨亚威.产胞外多糖乳酸菌的益生特性及其对成熟干酪品质的影响[D].北京:北京工商大学,2016.
[18]赵笑.酒曲中微生物产凝乳酶的特性及其应用研究[D].北京:北京工商大学,2015.
[19]YUN J J,KIELY L J,BARBANO D M,et al.Mozzarella cheese:impact of cooking temperature on chemical composition,proteolysis and functional properties 1[J].Journal of Dairy Science,1993,76(12):3664-3673.DOI:10.3168/jds.S0022-0302(93)77708-5.
[20]FORTIN M H,CHAMPAGNE C P,ST-GELAIS D,et al.Effect of time of inoculation,starter addition,oxygen level and salting on the viability of probiotic cultures during Cheddar cheese production[J].International Dairy Journal,2011,21(2):75-82.DOI:10.1016/j.idairyj.2010.09.007.
[21]DIEZHANDINO I,FERNANDEZ D,GONZALEZ L,et al.Microbiological,physico-chemical and proteolytic changes in a Spanish blue cheese during ripening[J].Food Chemistry,2015,168:134-141.DOI:10.1016/j.foodchem.2014.07.039.
[22]BORSTING M W,OVIST K B,RASMUSSEN M,et al.Impact of selected coagulants and starters on primary proteolysis and amino acid release related to bitterness and structure of reduced-fat Cheddar cheese[J].Dairy Science&Technology,2012,92(5):593-612.
[23]O’MAHONY J A,LUCEY J A,MCSWEENEY P L H.Chymosinmediated proteolysis,calcium solubilization,and texture development during the ripening of Cheddar cheese[J].Journal of Dairy Science,2005,88(9):3101-3114.DOI:10.3168/jds.S0022-0302(05)72992-1.
[24]SOODAM K,ONG L,POWELL I B,et al.Effect of calcium chloride addition and draining pH on the microstructure and texture of full fat Cheddar cheese during ripening[J].Food Chemistry,2015,181:111-118.DOI:10.1016/j.foodchem.2015.01.135.
[25]ONG L,DAGASTINE R R,KENTISH S E,et al.The effect of pH at renneting on the microstructure,composition and texture of Cheddar cheese[J].Food Research International,2012,48(1):119-130.DOI:10.1016/j.foodres.2012.02.020.
[26]艾对,张富新,李延华,等.顶空固态微萃取-气质联用法分析羊乳和牛乳中挥发性化合物[J].食品与发酵工业,2014,40(10):189-193.
[27]WOLF I V,PEROTTI M C,ZALAZAR C A.Composition and volatile profiles of commercial Argentinean blue cheeses[J].Journal of the Science of Food and Agriculture,2011,91(2):385-393.DOI:10.1002/jsfa.4198.
[28]李良.乳酸乳球菌发酵稀奶油过程中甲基酮的合成代谢调控[D].哈尔滨:哈尔滨工业大学,2015.