甲醇芽孢杆菌凝乳酶对马苏里拉干酪加工特性的影响
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摘要
为了探究甲醇芽孢杆菌(Bacillus methanolicus)凝乳酶在马苏里拉干酪加工中的应用,分别以使用甲醇芽孢杆菌凝乳酶、混合酶制剂(含质量分数10%甲醇芽孢杆菌凝乳酶和90%商品凝乳酶)制作的马苏里拉干酪作为实验组,以商品凝乳酶干酪作为对照组,测定不同组别干酪成熟期间的蛋白水解特性、质构、风味和微观结构变化,研究甲醇芽孢杆菌凝乳酶对马苏里拉干酪加工特性的影响。结果表明,实验组干酪在成熟过程中pH值(4.6～5.3)、微生物数量(8.80～9.68(lg(CFU/g)))与对照组无显著差异(P>0.05);实验组干酪水分质量分数(混合酶干酪为(43.21±1.17)%、甲醇芽孢杆菌凝乳酶干酪为(46.15±0.94)%)均显著高于对照组((41.08±1.04)%),得率(混合酶干酪为(9.27±0.17)%、甲醇芽孢杆菌凝乳酶干酪为(9.46±0.16)%)也显著高于对照组((8.98±0.13)%)(P<0.05);且实验组干酪的蛋白水解特性(pH 4.6时可溶性蛋白、酪蛋白水解程度和游离氨基酸质量分数)以及风味物质种类和相对含量等指标也优于对照组干酪。但是实验组中甲醇芽孢杆菌凝乳酶干酪保形性相对欠佳,感官评定得分偏低,混合酶干酪与对照组质构及感官基本得分一致,因此甲醇芽孢杆菌凝乳酶可以作为商品酶的部分代替品应用于干酪的生产中。
In order to explore the application of milk clotting enzyme(MCE) from Bacillus methanolicus in Mozzarella cheese making, single B. methanolicus MCE and an enzyme mixture of 10% B. methanolicus MCE and 90% commercial rennet were used separately in the preparation of Mozzarella cheese, while that made with commercial rennet was used as the control. The proteolytic properties, texture, flavor and microstructure of the cheeses were determined during the ripening process. Results showed that there was no significant difference in pH(4.6–5.3) or microbial count(8.80–9.68(lg(CFU/g)))between the three groups(P > 0.05). The moisture contents and cheese yields of the experimental groups((43.21 ± 1.17)% and((9.27 ± 0.17)% for single B. methanolicus MCE, and(46.15 ± 0.94)% and(9.46 ± 0.16)% for the enzyme mixture, respectively) were both significantly higher than those of the control group((41.08 ± 1.04)% moisture content and(8.98 ± 0.13)% cheese yield)(P < 0.05). Moreover, the experimental cheeses were also better than the control in terms of the proteolytic properties(the content of pH 4.6 soluble protein, the degree of casein hydrolysis and the content of free amino acids) and the types and contents of flavor substances. However, the cheese made with single B. methanolicus MCE had relatively poor shape-preserving property together with low sensory score. The cheese made with the enzyme mixture was comparable to the control in terms of texture and sensory scores, indicating that MCE from B. methanolicus could be used as a partial substitute for commercial rennet in cheese production.
In order to explore the application of milk clotting enzyme(MCE) from Bacillus methanolicus in Mozzarella cheese making, single B. methanolicus MCE and an enzyme mixture of 10% B. methanolicus MCE and 90% commercial rennet were used separately in the preparation of Mozzarella cheese, while that made with commercial rennet was used as the control. The proteolytic properties, texture, flavor and microstructure of the cheeses were determined during the ripening process. Results showed that there was no significant difference in pH(4.6–5.3) or microbial count(8.80–9.68(lg(CFU/g)))between the three groups(P > 0.05). The moisture contents and cheese yields of the experimental groups((43.21 ± 1.17)% and((9.27 ± 0.17)% for single B. methanolicus MCE, and(46.15 ± 0.94)% and(9.46 ± 0.16)% for the enzyme mixture, respectively) were both significantly higher than those of the control group((41.08 ± 1.04)% moisture content and(8.98 ± 0.13)% cheese yield)(P < 0.05). Moreover, the experimental cheeses were also better than the control in terms of the proteolytic properties(the content of pH 4.6 soluble protein, the degree of casein hydrolysis and the content of free amino acids) and the types and contents of flavor substances. However, the cheese made with single B. methanolicus MCE had relatively poor shape-preserving property together with low sensory score. The cheese made with the enzyme mixture was comparable to the control in terms of texture and sensory scores, indicating that MCE from B. methanolicus could be used as a partial substitute for commercial rennet in cheese production.
引文
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[11]郑远荣,刘振民,莫蓓红,等.切达奶酪成熟过程中微观结构变化及其对质构的影响[J].中国农业科学,2012,45(3):503-508.DOI:10.3864/j.issn.0578-1752.2012.03.012.
[12]李柳,郑喆,赵笑,等.黄酒麦曲中产凝乳酶菌株的分离鉴定[J].中国乳品工业,2017,45(11):4-10;18.DOI:10.3969/j.issn.1001-2230.2017.11.001.
[13]小文.《乳制品感官质量评鉴细则》正式发布[J].食品科学,2005,31(1):210.DOI:10.3321/j.issn:0253-990X.2005.03.040.
[14]JANA A H,TAGALPALLEWAR G P.Functional properties of Mozzarella cheese for its end use application[J].Journal of Food Science&Technology,2017,54(12):3766-3778.DOI:10.1007/s13197-017-2886-z.
[15]郝晓娜,张健,杨亚威,等.益生性植物乳杆菌对切达干酪挥发性风味形成的影响[J].食品科学,2018,39(4):49-58.DOI:10.7506/spkx1002-6630-201804009.
[16]VOIGT D D,CHEVALIER F,QIAN M C,et al.Effect of highpressure treatment on microbiology,proteolysis,lipolysis and levels of flavour compounds in mature blue-veined cheese[J].Innovative Food Science and Emerging Technologies,2010,11(1):68-77.DOI:10.1016/j.ifset.2009.10.009.
[17]梅俊.西藏开菲尔干酪的风味变化及可食膜包装研究[D].上海:上海交通大学,2015:1-129.
[18]陈俊亮,田芬,霍贵成,等.乳酸乳球菌对切达干酪成熟过程中质构和风味的影响[J].食品科学,2013,34(21):163-167.DOI:10.7506/spkx1002-6630-201321034.
[19]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.
[20]VINDEROLA G,PROSELLO W,MOLINARI F,et al.Growth of Lactobacillus paracasei A13 in Argentinian probiotic cheese and its impact on the characteristics of the product[J].International Journal of Food Microbiology,2009,135(2):171-174.DOI:10.1016/j.ijfoodmicro.2009.08.021.
[21]SULEJMANI E,HAYALOGLU A A.Influence of curd heating on proteolysis and volatiles of Kashkaval cheese[J].Food Chemistry,2016,211:160-170.DOI:10.1016/j.foodchem.2016.05.054.
[22]刘坚,牟光庆.奶酪风味物质形成机理的研究[J].食品研究与开发,2007,28(11):158-161.DOI:10.3969/j.issn.1005-6521.2007.11.049.
[23]GARCíA V,ROVIRA S,BOUTOIAL K,et al.Physicochemical,microbiological,textural and sensory changes during the ripening of pasteurised goat milk cheese made with plant coagulant[J].International Journal of Dairy Technology,2016,69(1):96-102.DOI:10.1111/1471-0307.12225.
[24]徐辉艳.木瓜蛋白酶对Mozzarella干酪成熟过程中蛋白降解及微观结构的影响[J].应用化工,2017,46(5):1020-1023.DOI:10.16581/j.cnki.issn1671-3206.2017.05.045.
[25]RADELJEVI B,NATA M,ANTUNAC N,et al.Influence of starter culture on total free aminoacids concentration during ripening of Krk cheese[J].Mljekarstvo,2013,63(1):533-538.DOI:10.7205/MILMED-D-14-00212.
[26]BERESFORD T,WILLIAMS A.The microbiology of cheese ripening[J].Cheese Chemistry Physics&Microbiology,2004,1(4):287-317.DOI:10.1016/S1874-558X(04)80071-X.
[27]B?RSTING M W,QVIST 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 and Technology,2012,92(5):593-612.DOI:10.1007/s13594-012-0080-7.
[28]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.
[29]ZHENG Y R,LIU Z M,BEI-HONG M O,et al.Effects of microstructure change on the texture of Cheddar cheese during ripening[J].Scientia Agricultura Sinica,2012,45(3):503-508.
[30]OLUK A C,GüVEN M,HAYALOGLU A A.Proteolysis texture and microstructure of low-fat Tulum cheese affected by exopolysaccharideproducing cultures during ripening[J].International Journal of Food Science&Technology,2014,49(2):435-443.DOI:10.1111/ijfs.12320.
[31]陈臣,段建平.原辅料对再制马苏里拉干酪品质特性的影响[J].食品科学,2017,38(1):182-187.DOI:10.7506/spkx1002-6630-201701030.
[32]ANDERSEN L,ANDERSEN L T.Flavour compounds and sensory characteristics of cheese powders made from matured cheeses[J].International Dairy Journal,2013,30(1):19-28.DOI:10.1016/j.idairyj.2012.11.002.
[2]NGUYEN H,ONG L,LOPEZ C,et al.Microstructure and physicochemical properties reveal differences between high moisture buffalo and bovine Mozzarella cheeses[J].Food Research International,2017,102:458-467.DOI:10.1016/j.foodres.2017.09.032.
[3]BERTA M,MUSKENS E,SCHUSTER E,et al.Rheology of natural and imitation Mozzarella cheese at conditions relevant to pizza baking[J].International Dairy Journal,2016,57:34-38.DOI:10.1016/j.idairyj.2016.02.038.
[4]MCSWEENEY P L H.Biochemistry of cheese ripening[J].International Journal of Dairy Technology,2010,57(2/3):127-144.DOI:10.1016/b978-0-12-374407-4.00080-7.
[5]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.
[6]HE X L,ZHANG W B,REN F Z,et al.Screening fermentation parameters of the milk-clotting enzyme produced by newly isolated Bacillus amyloliquefaciens D4 from the Tibetan Plateau in China[J].Annals of Microbiology,2012,62(1):357-365.DOI:10.1007/s13213-011-0270-1.
[7]NARWAL R K,BHUSHAN B,PAL A,et al.Optimization of upstream process parameters for enhanced production of thermostable milk clotting enzyme from Bacillus subtilis MTCC 10422[J].Journal of Food Process Engineering,2016,40(2):1-11.DOI:10.1111/jfpe.12356.
[8]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):34-36;47.DOI:10.3969/j.issn.1001-2230.2009.05.010.
[9]DING Z Y,WANG B F,OUYANG A,et al.Production and characterization of milk-clotting enzyme from Bacillus amyloliquefaciens JNU002 by submerged fermentation[J].European Food Research and Technology,2012,234(3):415-421.DOI:10.1007/s00217-011-1650-2.
[10]AHMED S A,WEHAIDY H R,IBRAHIM O A,et al.Novel milkclotting enzyme from Bacillus stearothermophilus as a coagulant in UF-white soft cheese[J].Biocatalysis&Agricultural Biotechnology,2016,7:241-249.DOI:10.1016/j.bcab.2016.06.011.
[11]郑远荣,刘振民,莫蓓红,等.切达奶酪成熟过程中微观结构变化及其对质构的影响[J].中国农业科学,2012,45(3):503-508.DOI:10.3864/j.issn.0578-1752.2012.03.012.
[12]李柳,郑喆,赵笑,等.黄酒麦曲中产凝乳酶菌株的分离鉴定[J].中国乳品工业,2017,45(11):4-10;18.DOI:10.3969/j.issn.1001-2230.2017.11.001.
[13]小文.《乳制品感官质量评鉴细则》正式发布[J].食品科学,2005,31(1):210.DOI:10.3321/j.issn:0253-990X.2005.03.040.
[14]JANA A H,TAGALPALLEWAR G P.Functional properties of Mozzarella cheese for its end use application[J].Journal of Food Science&Technology,2017,54(12):3766-3778.DOI:10.1007/s13197-017-2886-z.
[15]郝晓娜,张健,杨亚威,等.益生性植物乳杆菌对切达干酪挥发性风味形成的影响[J].食品科学,2018,39(4):49-58.DOI:10.7506/spkx1002-6630-201804009.
[16]VOIGT D D,CHEVALIER F,QIAN M C,et al.Effect of highpressure treatment on microbiology,proteolysis,lipolysis and levels of flavour compounds in mature blue-veined cheese[J].Innovative Food Science and Emerging Technologies,2010,11(1):68-77.DOI:10.1016/j.ifset.2009.10.009.
[17]梅俊.西藏开菲尔干酪的风味变化及可食膜包装研究[D].上海:上海交通大学,2015:1-129.
[18]陈俊亮,田芬,霍贵成,等.乳酸乳球菌对切达干酪成熟过程中质构和风味的影响[J].食品科学,2013,34(21):163-167.DOI:10.7506/spkx1002-6630-201321034.
[19]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.
[20]VINDEROLA G,PROSELLO W,MOLINARI F,et al.Growth of Lactobacillus paracasei A13 in Argentinian probiotic cheese and its impact on the characteristics of the product[J].International Journal of Food Microbiology,2009,135(2):171-174.DOI:10.1016/j.ijfoodmicro.2009.08.021.
[21]SULEJMANI E,HAYALOGLU A A.Influence of curd heating on proteolysis and volatiles of Kashkaval cheese[J].Food Chemistry,2016,211:160-170.DOI:10.1016/j.foodchem.2016.05.054.
[22]刘坚,牟光庆.奶酪风味物质形成机理的研究[J].食品研究与开发,2007,28(11):158-161.DOI:10.3969/j.issn.1005-6521.2007.11.049.
[23]GARCíA V,ROVIRA S,BOUTOIAL K,et al.Physicochemical,microbiological,textural and sensory changes during the ripening of pasteurised goat milk cheese made with plant coagulant[J].International Journal of Dairy Technology,2016,69(1):96-102.DOI:10.1111/1471-0307.12225.
[24]徐辉艳.木瓜蛋白酶对Mozzarella干酪成熟过程中蛋白降解及微观结构的影响[J].应用化工,2017,46(5):1020-1023.DOI:10.16581/j.cnki.issn1671-3206.2017.05.045.
[25]RADELJEVI B,NATA M,ANTUNAC N,et al.Influence of starter culture on total free aminoacids concentration during ripening of Krk cheese[J].Mljekarstvo,2013,63(1):533-538.DOI:10.7205/MILMED-D-14-00212.
[26]BERESFORD T,WILLIAMS A.The microbiology of cheese ripening[J].Cheese Chemistry Physics&Microbiology,2004,1(4):287-317.DOI:10.1016/S1874-558X(04)80071-X.
[27]B?RSTING M W,QVIST 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 and Technology,2012,92(5):593-612.DOI:10.1007/s13594-012-0080-7.
[28]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.
[29]ZHENG Y R,LIU Z M,BEI-HONG M O,et al.Effects of microstructure change on the texture of Cheddar cheese during ripening[J].Scientia Agricultura Sinica,2012,45(3):503-508.
[30]OLUK A C,GüVEN M,HAYALOGLU A A.Proteolysis texture and microstructure of low-fat Tulum cheese affected by exopolysaccharideproducing cultures during ripening[J].International Journal of Food Science&Technology,2014,49(2):435-443.DOI:10.1111/ijfs.12320.
[31]陈臣,段建平.原辅料对再制马苏里拉干酪品质特性的影响[J].食品科学,2017,38(1):182-187.DOI:10.7506/spkx1002-6630-201701030.
[32]ANDERSEN L,ANDERSEN L T.Flavour compounds and sensory characteristics of cheese powders made from matured cheeses[J].International Dairy Journal,2013,30(1):19-28.DOI:10.1016/j.idairyj.2012.11.002.