发酵剂及猕猴桃蛋白酶促进马肉发酵香肠蛋白质的降解
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摘要
为了提高马肉发酵香肠的品质,添加发酵剂和猕猴桃蛋白酶,促进其发酵成熟和肌肉蛋白质的降解。以新鲜马肉和马脂肪为原料,分别设计三个试验组,一组为对照组,另两组为处理组;对照组不采取处理,而两个处理组添加2%的发酵剂和0.05%的猕猴桃蛋白酶,并在相应条件下进行发酵,检测其总氮、非蛋白氮等指标,并通过SDS-PAGE电泳分析其肌肉蛋白质的降解情况。结果表明,灌肠后(0 d)各组总氮含量为3.09 g/100 g左右,而加工结束后(28 d),对照组(CK组)、发酵组(A组)、猕猴桃蛋白酶组(B组)总氮含量分别是3.41 g/100 g、3.85 g/100 g及4.15 g/100 g,说明总氮含量明显上升,并且发酵剂和猕猴桃蛋白酶的影响显著;28 d CK组、A组、B组非蛋白氮含量分别为0.42 g/100 g、0.52 g/100 g、0.65 g/100 g,与0 d非蛋白氮含量0.22 g/100 g相比,CK组上升1.9倍,A组上升2.4倍,B组上升2.9倍;SDS-PAGE电泳结果表明,在整个加工过程中A组与B组肌浆蛋白逐渐发生降解,大分子条带发生了明显的变化,发酵剂对马肉发酵香肠肌浆蛋白和肌原纤维蛋白的降解有显著的促进作用,猕猴桃蛋白酶对肌浆蛋白的影响显著,并比发酵剂大,但对肌原纤维蛋白的作用不大。
In order to improve the quality of horse meat fermented sausage, starter and actinidin were added to promote the fermentation mature and degradation of muscle protein. Fresh horse meat and horse fat were chose as raw materials. Three test groups were designed. One was the control group and the other two groups were the treatment group. The two treatment groups added 2% starter and 0.05% actinidin, and fermented under the corresponding conditions. the contents of total nitrogen, non-protein nitrogen were detected. The degradation of muscle protein was analyzed by SDS-PAGE. Results showed that the total nitrogen content of each group(0 d) was about 3.09 g/100 g. After processing(28 d), the total nitrogen content of control group(CK group), fermentation group(group A) and actinidin group(group B) were 3.41 g/100 g,3.85 g/100 g and 4.15 g/100 g, respectively, indicating that the total nitrogen content increased significantly. The effects of starter and actinidin on protein degradation were significant. The non-protein nitrogen contents of the CK group, A group, B group on 28 d were 0.42 g/100 g, 0.52 g/100 g, 0.65 g/100 g respectively. Compared with 0 d(0.22 g/100 g), the non-protein nitrogen content of CK group increased by 1.9 times,group A increased by 2.4 times, group B increased by 2.9 times. Results of SDS-PAGE electrophoresis showed that the sarcoplasmic protein of group A and group B gradually degraded during the whole processing, and the macromolecular bands changed significantly. The starter significantly promoted the degradation of sarcoplasmic protein and myofibrillar protein of horse meat fermented sausage. Actinidin can also promote the sarcoplasmic protein degradation and have better effect than starter, but it has little effect on myofibrillar protein.
In order to improve the quality of horse meat fermented sausage, starter and actinidin were added to promote the fermentation mature and degradation of muscle protein. Fresh horse meat and horse fat were chose as raw materials. Three test groups were designed. One was the control group and the other two groups were the treatment group. The two treatment groups added 2% starter and 0.05% actinidin, and fermented under the corresponding conditions. the contents of total nitrogen, non-protein nitrogen were detected. The degradation of muscle protein was analyzed by SDS-PAGE. Results showed that the total nitrogen content of each group(0 d) was about 3.09 g/100 g. After processing(28 d), the total nitrogen content of control group(CK group), fermentation group(group A) and actinidin group(group B) were 3.41 g/100 g,3.85 g/100 g and 4.15 g/100 g, respectively, indicating that the total nitrogen content increased significantly. The effects of starter and actinidin on protein degradation were significant. The non-protein nitrogen contents of the CK group, A group, B group on 28 d were 0.42 g/100 g, 0.52 g/100 g, 0.65 g/100 g respectively. Compared with 0 d(0.22 g/100 g), the non-protein nitrogen content of CK group increased by 1.9 times,group A increased by 2.4 times, group B increased by 2.9 times. Results of SDS-PAGE electrophoresis showed that the sarcoplasmic protein of group A and group B gradually degraded during the whole processing, and the macromolecular bands changed significantly. The starter significantly promoted the degradation of sarcoplasmic protein and myofibrillar protein of horse meat fermented sausage. Actinidin can also promote the sarcoplasmic protein degradation and have better effect than starter, but it has little effect on myofibrillar protein.
引文
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[11]王海丽.软枣猕猴桃蛋白酶的提取分离与嫩化牛肉效果的研究[D].延吉:延边大学,2013WANG Hai-li.Study on the extraction and separation of soft kiwifruit protease and tenderized beef[D].Yanji:Yanbian University,2013
[12]Han J,Morton J D,Bekhit A E,et al.Pre-rigor infusion with kiwifruit juice improves lamb tenderness[J].Meat Science,2009,82(3):324-330
[13]Aminlari M,Shekarforoush S S,Gheisari H R,et al.Effect of actinidin on the protein solubility,water holding capacity,texture,electrophoretic pattern of beef,and on the quality attributes of a sausage product[J].Journal of Food Science,2009,74(3):221-226
[14]Hashim M M,Mingsheng D,Iqbal M F,et al.Ginger rhizome as a potential source of milk coagulating cysteine protease[J].Phytochemistry,2011,72(6):458-464
[15]Kamphuis I G,Drenth J,Baker E N,et al.Thiol proteases:Comparative studies based on the high-resolution structures of papain and actinidin,and on amino acid sequence information for cathepsins B and H,and stem bromelain[J].Journal of Molecular Biology,1985,182(2):317-329
[16]王恺.三种乳酸菌发酵剂在发酵香肠中的应用研究[D].南京:南京农业大学,2013WANG Kai.Application of three lactic acid bacteria starter in fermented sausage[D].Nanjing:Nanjing Agricultural University,2013
[17]张雪梅.四川香肠生产过程中理化特性,微生物特性及产香葡萄球菌的筛选与应用[D].雅安:四川农业大学,2010ZHANG Xue-mei.Physicochemical properties,microbial characteristics and screening and application of Staphylococcus aureus in Sichuan sausage production[D].Ya'an:Sichuan Agricultural University,2010
[18]朱健辉.不同等级金华火腿蛋白质和脂肪水解产物与挥发性风味物质分析[D].南京:南京农业大学,2005ZHU Jian-hui.Analysis of protein and fat hydrolysate and hair flavor substances of different grades of Jinhua ham[D].Nanjing:Nanjing Agricultural University,2005
[19]Molina I,Toldra F.Detection of proteolytic activity in microorganisms isolated from dry-cured ham[J].Journal of Food Science,1992,57(6):1308-1310
[20]Fadda S,Sanz Y,Vignolo G,et al.Characterization of muscle sarcoplasmic and myofibrillar protein hydrolysis caused by Lactobacillus plantarum[J].Appl Environ Microbiol,1999,65(8):3540-3546
[21]Aminlari M,Shekarforoush S S,Gheisari H R,et al.Effect of actinidin on the protein solubility,water holding capacity,texture,electrophoretic pattern of beef,and on the quality attributes of a sausage product[J].Journal of Food Science,2009,74(3):C221-C226
[22]吕舒.四川香肠加工贮藏过程中表层,中层,内层的理化特性及微生物的变化[D].雅安:四川农业大学,2014LYU Shu.Physicochemical properties and microbial changes in the surface,middle and inner layers of Sichuan sausage during processing and storage[D].Ya'an:Sichuan Agricultural University,2014
[23]孙霞,巩洋,杨勇,等.接种混合发酵剂生产的低酸度川味香肠加工过程中的蛋白质降解[J].食品工业科技,2015,36(13):66-71SUN Xia,GONG Yang,YANG Yong,et al.Protein degradation during processing of low acidity Sichuan sausages inoculated with mixed starter[J].Food Science and Technology,2015,36(13):66-71
[24]Soriano A,Garcia R A,Gomez E,et al.Lipolysis,proteolysis,physicochemical and sensory characteristics of different types of Spanish ostrich salchichon[J].Meat Science,2007,75(4):661-668
[25]Díaz O,Fernández M,Fernando G D G D,et al.Effect of the addition of papain on the dry fermented sausage proteolysis[J].Journal of the Science of Food&Agriculture,2015,71(1):13-21
[26]韩鲜娜.新疆熏马肠强化接种与内源酶调控技术研究[D].石河子市:石河子大学,2013HAN Xian-na.Study on intensive inoculation and endogenous enzyme control techniques of smoked horse in Xinjiang[D].Shihezi:Shihezi University,2013
[27]Zhao G M,Zhou G H,Wang Y L,et al.Time-related changes in cathepsin B and L activities during processing of Jinhua ham as a function of p H,salt and temperature[J].Meat Science,2005,70(2):381-388
[28]Hughes M C,Kerry J P,Arendt E K,et al.Characterization of proteolysis during the ripening of semi-dry fermented sausages[J].Meat Science,2002,62(2):205-216
[29]Fadda S,Sanz Y,Vignolo G,et al.Characterization of muscle sarcoplasmic and myofibrillar protein hydrolysis caused by Lactobacillus plantarum[J].Appl Environ Microbiol,1999,65(8):3540-3546
[30]Candogan K,Wardlaw F B,Acton J C,et al.Effect of starter culture on proteolytic changes during processing of fermented beef sausages[J].Food Chemistry,2009,116(3):731-737
[2]Yang H,Zhang L,Li-Zhen M A,et al.Effect of exogenous enzymes on shortening ripening period of fermented mutton sausage[J].Food Science,2010,15:81-86
[3]Ansorena D,Zapelena M J,Astiasarán I,et al.Simultaneous addition of palatase M and protease P to a dry fermented sausage(Chorizo de Pamplona)elaboration:Effect over peptidic and Lipid fractions[J].Meat Science,1998,50(1):37-44
[4]段艳.乳酸菌的筛选及其对羊肉干发酵香肠品质特性的影响[D].呼和浩特市:内蒙古农业大学,2013DUAN Yan.Screening of lactic acid bacteria and its effect on quality characteristics of mutton fermented sausage[D].Hohhot:Inner Mongolia Agricultural University,2013
[5]孔令明,李芳,张文,等.不同电刺激处理时间对宰后马肉成熟过程中嫩度的影响[J].食品科学,2018,39(9):76-81KONG Ling-ming,LI Fang,ZHANG Wen,et al.Effect of electrical stimulation on horse meat tenderness during postmortem aging[J].Food Science,2018,39(9):76-81
[6]Alagi?,Koza?inski L,Filipovi?I,et al.Microbial changes during ripening of fermented horse meat sausages[J].Meso Prvi Hrvatski?asopis O Mesu,2008,10(3):200-203
[7]Badiani A,Nanni N,Gatta P P,et al.Nutrient profile of horsemeat[J].Journal of Food Composition&Analysis,1997,10(3):254-269
[8]赵立男,热夏提达·吾来提,杨随霞,等.马肉不同部位的品质特性分析[J].肉类研究,2013,27(7):35-39ZHAO Li-nan,RE Xia-ti-da·WU Lai-ti,YANG Sui-xia,et al.Analysis of quality characteristics of different parts of horse meat[J].Meat Research,2013,27(7):35-39
[9]杨红,徐明华.1961-2010年世界马肉生产与效率分析[J].经济研究导刊,2012,29:31-33YANG Hong,XU Ming-hua.Analysis of world horse meat production and efficiency from 1961 to 2010[J].Economic Research Guide,2012,29:31-33
[10]Katsaros G I,Tavantzis G,Taoukis P S,et al.Production of novel dairy products using actinidin and high pressure as enzyme activity regulator[J].Innovative Food Science&Emerging Technologies,2010,11(1):47-51
[11]王海丽.软枣猕猴桃蛋白酶的提取分离与嫩化牛肉效果的研究[D].延吉:延边大学,2013WANG Hai-li.Study on the extraction and separation of soft kiwifruit protease and tenderized beef[D].Yanji:Yanbian University,2013
[12]Han J,Morton J D,Bekhit A E,et al.Pre-rigor infusion with kiwifruit juice improves lamb tenderness[J].Meat Science,2009,82(3):324-330
[13]Aminlari M,Shekarforoush S S,Gheisari H R,et al.Effect of actinidin on the protein solubility,water holding capacity,texture,electrophoretic pattern of beef,and on the quality attributes of a sausage product[J].Journal of Food Science,2009,74(3):221-226
[14]Hashim M M,Mingsheng D,Iqbal M F,et al.Ginger rhizome as a potential source of milk coagulating cysteine protease[J].Phytochemistry,2011,72(6):458-464
[15]Kamphuis I G,Drenth J,Baker E N,et al.Thiol proteases:Comparative studies based on the high-resolution structures of papain and actinidin,and on amino acid sequence information for cathepsins B and H,and stem bromelain[J].Journal of Molecular Biology,1985,182(2):317-329
[16]王恺.三种乳酸菌发酵剂在发酵香肠中的应用研究[D].南京:南京农业大学,2013WANG Kai.Application of three lactic acid bacteria starter in fermented sausage[D].Nanjing:Nanjing Agricultural University,2013
[17]张雪梅.四川香肠生产过程中理化特性,微生物特性及产香葡萄球菌的筛选与应用[D].雅安:四川农业大学,2010ZHANG Xue-mei.Physicochemical properties,microbial characteristics and screening and application of Staphylococcus aureus in Sichuan sausage production[D].Ya'an:Sichuan Agricultural University,2010
[18]朱健辉.不同等级金华火腿蛋白质和脂肪水解产物与挥发性风味物质分析[D].南京:南京农业大学,2005ZHU Jian-hui.Analysis of protein and fat hydrolysate and hair flavor substances of different grades of Jinhua ham[D].Nanjing:Nanjing Agricultural University,2005
[19]Molina I,Toldra F.Detection of proteolytic activity in microorganisms isolated from dry-cured ham[J].Journal of Food Science,1992,57(6):1308-1310
[20]Fadda S,Sanz Y,Vignolo G,et al.Characterization of muscle sarcoplasmic and myofibrillar protein hydrolysis caused by Lactobacillus plantarum[J].Appl Environ Microbiol,1999,65(8):3540-3546
[21]Aminlari M,Shekarforoush S S,Gheisari H R,et al.Effect of actinidin on the protein solubility,water holding capacity,texture,electrophoretic pattern of beef,and on the quality attributes of a sausage product[J].Journal of Food Science,2009,74(3):C221-C226
[22]吕舒.四川香肠加工贮藏过程中表层,中层,内层的理化特性及微生物的变化[D].雅安:四川农业大学,2014LYU Shu.Physicochemical properties and microbial changes in the surface,middle and inner layers of Sichuan sausage during processing and storage[D].Ya'an:Sichuan Agricultural University,2014
[23]孙霞,巩洋,杨勇,等.接种混合发酵剂生产的低酸度川味香肠加工过程中的蛋白质降解[J].食品工业科技,2015,36(13):66-71SUN Xia,GONG Yang,YANG Yong,et al.Protein degradation during processing of low acidity Sichuan sausages inoculated with mixed starter[J].Food Science and Technology,2015,36(13):66-71
[24]Soriano A,Garcia R A,Gomez E,et al.Lipolysis,proteolysis,physicochemical and sensory characteristics of different types of Spanish ostrich salchichon[J].Meat Science,2007,75(4):661-668
[25]Díaz O,Fernández M,Fernando G D G D,et al.Effect of the addition of papain on the dry fermented sausage proteolysis[J].Journal of the Science of Food&Agriculture,2015,71(1):13-21
[26]韩鲜娜.新疆熏马肠强化接种与内源酶调控技术研究[D].石河子市:石河子大学,2013HAN Xian-na.Study on intensive inoculation and endogenous enzyme control techniques of smoked horse in Xinjiang[D].Shihezi:Shihezi University,2013
[27]Zhao G M,Zhou G H,Wang Y L,et al.Time-related changes in cathepsin B and L activities during processing of Jinhua ham as a function of p H,salt and temperature[J].Meat Science,2005,70(2):381-388
[28]Hughes M C,Kerry J P,Arendt E K,et al.Characterization of proteolysis during the ripening of semi-dry fermented sausages[J].Meat Science,2002,62(2):205-216
[29]Fadda S,Sanz Y,Vignolo G,et al.Characterization of muscle sarcoplasmic and myofibrillar protein hydrolysis caused by Lactobacillus plantarum[J].Appl Environ Microbiol,1999,65(8):3540-3546
[30]Candogan K,Wardlaw F B,Acton J C,et al.Effect of starter culture on proteolytic changes during processing of fermented beef sausages[J].Food Chemistry,2009,116(3):731-737