贻贝蛋白酶解体系稳定性影响因素及其机制
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摘要
贻贝酶解液(HMP)中蛋白质稳定性是决定其相关产品品质的关键因素。本试验中采用凯氏定氮、SDSPAGE技术、Bradford法、激光粒度分析法等方法研究了不同因素对HMP蛋白质稳定性的影响,分析了蛋白浓度、p H值对不同酶解条件下HMP中蛋白沉淀率和体系分子粒度变化的相关性,并以此衡量体系的稳定性。结果发现,蛋白浓度、pH值对HMP蛋白酶解体系分子的粒度和稳定性均有较大影响。碱性体系比酸性体系更有利于HMP的稳定;随着蛋白浓度的增加蛋白质稳定性降低;粒度分布均匀的HMP比较稳定。本研究为HMP稳定性的研究提供了一定的理论依据,为贻贝多肽功能性饮料等产品的开发提供技术参考。
The stability of hydrolysate of protein from Mytilus edulis by enzymatic hydrolysis plays an important role in keeping the quality of products made from these peptides. In the present study, the laser particle size analysis, SDSPAGE, Bradford and Kjeldahl method were used to investigate the influence factors on stability of HMP. The relative protein yield and the change of particle size of HMP were compared under various conditions, including a range of protein concentration, p H values, and different enzymatic hydrolysis parameters, was used to evaluate the stability of HMP.Results showed that protein concentration, p H, and particle size played an important role in the stability of HMP. The protein stability in alkaline system was better than that in acidic system. With the increase of protein concentration, protein stability decreased. Moreover, the HMP with uniform particle size distribution would be relative more stable. The study provided a certain theoretical basis for the study on protein stability of HMP, and provided technical references for development of functional beverage product made from mytilus edulis polypeptides.
The stability of hydrolysate of protein from Mytilus edulis by enzymatic hydrolysis plays an important role in keeping the quality of products made from these peptides. In the present study, the laser particle size analysis, SDSPAGE, Bradford and Kjeldahl method were used to investigate the influence factors on stability of HMP. The relative protein yield and the change of particle size of HMP were compared under various conditions, including a range of protein concentration, p H values, and different enzymatic hydrolysis parameters, was used to evaluate the stability of HMP.Results showed that protein concentration, p H, and particle size played an important role in the stability of HMP. The protein stability in alkaline system was better than that in acidic system. With the increase of protein concentration, protein stability decreased. Moreover, the HMP with uniform particle size distribution would be relative more stable. The study provided a certain theoretical basis for the study on protein stability of HMP, and provided technical references for development of functional beverage product made from mytilus edulis polypeptides.
引文
[1]农业部渔业局.2016年中国渔业统计年鉴[M].北京:中国农业出版社,2016:29.
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[3]张艳萍,戴志远,张虹.贻贝中ACE抑制活性肽的酶解制备及表征[J].中国食品学报,2011,11(1):51-59.
[4]ASOKAN P K,MOHAMED K S,SASIKUMAR GMussel farming methods and the prospects and problems in India[J].Course Manual,2016,18(3):41-46.
[5]周越.贻贝肽与贻贝多糖对衰老的干预作用及其机制[D].镇江:江苏大学,2013.
[6]BUI L M,BIERER T L.Influence of green lipped mussels(Perna canaliculus)in alleviating signs of arthritis in dogs[J].Veterinary Therapeutics,2003,2(2):397-407.
[7]TAYLOR A G,SAVAGE C.Fatty acid composition of New Zealand green-lipped mussels,Perna canaliculus:Implications for harvesting for n-3 extracts[J].Aquaculture,2006,261(1):430-439.
[8]OROZCO Z G A,SUMBING J G,LEBATA-RAMOS M J H,et al.Apparent digestibility coefficient of nutrients from shrimp,mussel,diatom and seaweed by juvenile Holothuria scabra,Jaeger[J]Aquaculture Research,2014,45(7):1153-1163.
[9]MITTA G,HUBERT F,NOEL T.et al.Myticin,a novel cysteine-rich antimicrobial peptide isolated from haemocytes and plasma of the mussel Mytilus galloprovincialis[J].European Journal Biochemistry,1999,265(1):71-78.
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[11]何建瑜,赵荣涛,刘慧慧.舟山海域厚壳贻贝软体部分营养成分分析与评价[J].南方水产科学,2012,8(4):37-42.
[12]BARRETT D G,FULLENKAMP D E,HE L,et al.p H‐Based regulation of hydrogel mechanical properties through mussel‐inspired chemistry and processing[J].Advanced functional materials,2013,23(9):1111-1119.
[13]NGO D,WIJESEKARA I,VO T,et al.Marine food-derived functional ingredients as potential antioxidants in the food industry:An overview[J].Food Res Int,2011,44(2):523-529.
[14]段伟文,罗伟,段振华,等.贻贝的加工利用研究进展[J].渔业现代化,2013,40(3):51-55.
[15]李芳,赵广林,蒋小锋,等.鲍鱼果蛋白饮料的工艺优化及其稳定性研究[J].食品研究与开发,2015,36(4):81-84.
[16]肖岚,李诚,付刚,等.5种蛋白酶对猪皮胶原蛋白水解效果的比较研究[J].食品研究与开发,2015,36(17):10-14.
[17]THUY C X,LAM T B.Biochemical and functional properties of fish protein isolate(FPI)from Pangasius hypophthalmus byproducts as influenced by time and degree of hydrolysis(DH)[J].International Food Research Journal,2015,22(1):2080-2085.
[18]FENG J,DAI Z,ZHANG Y,et al.Alteration of gene expression profile in kidney of spontaneously hypertensive rats treated with protein hydrolysate of blue mussel(Mytilus edulis)by DNA microarray analysis[J].Plos One,2015,10(10):e0142016-.
[19]YE L,LIAO Y,SUN W,et al.Effect of protein oxidation on the stability of peanut beverage[J].CyTA-Journal of Food,2015,13(1):49-55.
[20]郑和堂.激光光散射法测定百树菊酯聚合物纳米粒子的粒度[J].中华卫生杀虫药械,2010,26(2):102-104.
[21]柳荫,吴凤智,陈龙,等.考马斯亮蓝法测定核桃水溶性蛋白含量的研究[J].中国酿造,2013,32(12):131-133.
[22]QI H,NIU F,YI M,et al.Research on aerosol filtration in containment vessel by MEMS inertial impactor[J].Nuclear Science and Technology,2015,3(1):22-28.
[23]刘爽,袁芳,高彦祥.植物蛋白乳液稳定性研究进展[J].中国食品添加剂,2014,8(1):165-171.
[24]LUCEY J A,TAMEHANA M,SINGH H,et al Stability of model acid milk beverage:Effect of pectin concentration,storage temperature and milk heat treatment[J].Journal of Texture Studies,2007,30(3):305-318.
[25]BANACH J C.Modified milk protein concentrates in high-protein nutrition bars[D].2016.
[26]SASIKUMAR R.Preparation of therapeutic RTSbeverage from aloe vera gel and aonla fruit juice and evaluation of storage stability[J].Journal of Dairying Foods&Home Sciences,2015,34(2):151.
[27]陈文芬,崔春,肖如武,等.酸性鱼蛋白饮料的稳定性研究[J].现代食品科技,2008,24(1):1-4.
[2]TU K,SHAO X F,CHEN L,et al.Effects of pre-storage hot air treatments on the post-harvest quality and blue mold control of‘Red Fuji’apple fruit[J].Acta Horticulturae,2006,34(3):793-798.
[3]张艳萍,戴志远,张虹.贻贝中ACE抑制活性肽的酶解制备及表征[J].中国食品学报,2011,11(1):51-59.
[4]ASOKAN P K,MOHAMED K S,SASIKUMAR GMussel farming methods and the prospects and problems in India[J].Course Manual,2016,18(3):41-46.
[5]周越.贻贝肽与贻贝多糖对衰老的干预作用及其机制[D].镇江:江苏大学,2013.
[6]BUI L M,BIERER T L.Influence of green lipped mussels(Perna canaliculus)in alleviating signs of arthritis in dogs[J].Veterinary Therapeutics,2003,2(2):397-407.
[7]TAYLOR A G,SAVAGE C.Fatty acid composition of New Zealand green-lipped mussels,Perna canaliculus:Implications for harvesting for n-3 extracts[J].Aquaculture,2006,261(1):430-439.
[8]OROZCO Z G A,SUMBING J G,LEBATA-RAMOS M J H,et al.Apparent digestibility coefficient of nutrients from shrimp,mussel,diatom and seaweed by juvenile Holothuria scabra,Jaeger[J]Aquaculture Research,2014,45(7):1153-1163.
[9]MITTA G,HUBERT F,NOEL T.et al.Myticin,a novel cysteine-rich antimicrobial peptide isolated from haemocytes and plasma of the mussel Mytilus galloprovincialis[J].European Journal Biochemistry,1999,265(1):71-78.
[10]JE J Y,PARK P J,BYUN H G,et al.Angiotensin I converting enzyme(ACE)inhibitory peptide derived from the sauce of fermented bluemussel Mytilus edulis[J].Bioresource Technology,2005,96(14):1624-1629.
[11]何建瑜,赵荣涛,刘慧慧.舟山海域厚壳贻贝软体部分营养成分分析与评价[J].南方水产科学,2012,8(4):37-42.
[12]BARRETT D G,FULLENKAMP D E,HE L,et al.p H‐Based regulation of hydrogel mechanical properties through mussel‐inspired chemistry and processing[J].Advanced functional materials,2013,23(9):1111-1119.
[13]NGO D,WIJESEKARA I,VO T,et al.Marine food-derived functional ingredients as potential antioxidants in the food industry:An overview[J].Food Res Int,2011,44(2):523-529.
[14]段伟文,罗伟,段振华,等.贻贝的加工利用研究进展[J].渔业现代化,2013,40(3):51-55.
[15]李芳,赵广林,蒋小锋,等.鲍鱼果蛋白饮料的工艺优化及其稳定性研究[J].食品研究与开发,2015,36(4):81-84.
[16]肖岚,李诚,付刚,等.5种蛋白酶对猪皮胶原蛋白水解效果的比较研究[J].食品研究与开发,2015,36(17):10-14.
[17]THUY C X,LAM T B.Biochemical and functional properties of fish protein isolate(FPI)from Pangasius hypophthalmus byproducts as influenced by time and degree of hydrolysis(DH)[J].International Food Research Journal,2015,22(1):2080-2085.
[18]FENG J,DAI Z,ZHANG Y,et al.Alteration of gene expression profile in kidney of spontaneously hypertensive rats treated with protein hydrolysate of blue mussel(Mytilus edulis)by DNA microarray analysis[J].Plos One,2015,10(10):e0142016-.
[19]YE L,LIAO Y,SUN W,et al.Effect of protein oxidation on the stability of peanut beverage[J].CyTA-Journal of Food,2015,13(1):49-55.
[20]郑和堂.激光光散射法测定百树菊酯聚合物纳米粒子的粒度[J].中华卫生杀虫药械,2010,26(2):102-104.
[21]柳荫,吴凤智,陈龙,等.考马斯亮蓝法测定核桃水溶性蛋白含量的研究[J].中国酿造,2013,32(12):131-133.
[22]QI H,NIU F,YI M,et al.Research on aerosol filtration in containment vessel by MEMS inertial impactor[J].Nuclear Science and Technology,2015,3(1):22-28.
[23]刘爽,袁芳,高彦祥.植物蛋白乳液稳定性研究进展[J].中国食品添加剂,2014,8(1):165-171.
[24]LUCEY J A,TAMEHANA M,SINGH H,et al Stability of model acid milk beverage:Effect of pectin concentration,storage temperature and milk heat treatment[J].Journal of Texture Studies,2007,30(3):305-318.
[25]BANACH J C.Modified milk protein concentrates in high-protein nutrition bars[D].2016.
[26]SASIKUMAR R.Preparation of therapeutic RTSbeverage from aloe vera gel and aonla fruit juice and evaluation of storage stability[J].Journal of Dairying Foods&Home Sciences,2015,34(2):151.
[27]陈文芬,崔春,肖如武,等.酸性鱼蛋白饮料的稳定性研究[J].现代食品科技,2008,24(1):1-4.