响应面法优化近江牡蛎多糖多肽联产的酶解工艺
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摘要
利用响应面法优化近江牡蛎(Ostrea rivularis)多糖多肽联产的酶解工艺条件。通过单因素实验,研究酶解时间、酶配比、料液比对水解度、多肽含量、酸性糖含量以及总糖含量的影响,在此基础上,应用响应面试验建立数学模型,以多肽含量、酸性糖含量和总糖含量为响应值,进行三因素三水平的响应面分析。结果表明,酶解法联产制备近江牡蛎多糖多肽的最佳工艺为时间2.0 h,添加配比为3∶1的alcalase和胰蛋白酶(总质量分数为0.8%),料液比1∶3.6 g/mL。此条件下进行重复试验,得到近江牡蛎酶解液中多肽含量(251.70±5.73) mg·50 g~(-1),酸性糖含量(8.17±0.29) mg·50 g~(-1),总糖含量(380.66±15.14) mg·50 g~(-1)。并按此工艺酶解近江牡蛎全脏器匀浆,得粗多糖21.29%±0.77%,含85.18%±1.00%总糖,20.02%±0.38%酸性糖以及10.84%±0.12%多肽;粗多肽得率为31.41%±0.47%,含84.03%±0.85%多肽,0.11%±0.01%酸性糖以及5.68%±0.15%总糖。与回归方程预测值相近,说明近江牡蛎多糖多肽联产工艺是可行的。
The optimal enzymolysis conditions of simultaneous extraction of polysaccharides and polypeptides were explored from Ostrea rivularis by response surface methodology.Single factors experiments were used to find out the effects of hydrolysis time,enzymolysis proportion and solid-liquid ratio on the degree of hydrolysis,polypeptide content,glycosaminoglycan content and total-sugar content.Then three mathematical models were set up by response surface experiment.With three response values including polypeptide content,glycosaminoglycan content and total-sugar content,the three-factor and three-level response surface analysis was set up. The results showed that the optimal process for the preparation of polysaccharides from Oujiang oysters by enzymatic hydrolysis was 2.0 h,adding 3 ∶ 1 alcalase and trypsin( total mass fraction of 0.8%),and the ratio of material to liquid was 1∶ 3.6 g/mL.Under these conditions,repeated experiments were carried out to determine that the content of polypeptides was( 251.70 ± 5.73) mg·50 g~(-1),the content of glycosaminoglycans was( 8.17 ± 0.29) mg·50 g~(-1)and the content of total-sugar was( 380.66 ± 15.14) mg·50 g~(-1). The yield of rough polysaccharides in whole organ of Ostrea rivularis was 21.29 ± 0.77%,which contained 85.15% ± 1.00% total-sugar,20.02% ± 0.38% glycosaminoglycans and 10.84% ±0.12% polypeptides.Otherwise the yield of rough polypeptides was 31.41% ± 0.47%,which contained 84.03% ± 0.85%polypeptides,0.11% ± 0.01% glycosaminoglycans and 5.68% ± 0.15% total-sugar.In one word,the co-production process for polysaccharides and polypeptides from Ostrea rivularis was feasible,basically the same as the predicted value.
The optimal enzymolysis conditions of simultaneous extraction of polysaccharides and polypeptides were explored from Ostrea rivularis by response surface methodology.Single factors experiments were used to find out the effects of hydrolysis time,enzymolysis proportion and solid-liquid ratio on the degree of hydrolysis,polypeptide content,glycosaminoglycan content and total-sugar content.Then three mathematical models were set up by response surface experiment.With three response values including polypeptide content,glycosaminoglycan content and total-sugar content,the three-factor and three-level response surface analysis was set up. The results showed that the optimal process for the preparation of polysaccharides from Oujiang oysters by enzymatic hydrolysis was 2.0 h,adding 3 ∶ 1 alcalase and trypsin( total mass fraction of 0.8%),and the ratio of material to liquid was 1∶ 3.6 g/mL.Under these conditions,repeated experiments were carried out to determine that the content of polypeptides was( 251.70 ± 5.73) mg·50 g~(-1),the content of glycosaminoglycans was( 8.17 ± 0.29) mg·50 g~(-1)and the content of total-sugar was( 380.66 ± 15.14) mg·50 g~(-1). The yield of rough polysaccharides in whole organ of Ostrea rivularis was 21.29 ± 0.77%,which contained 85.15% ± 1.00% total-sugar,20.02% ± 0.38% glycosaminoglycans and 10.84% ±0.12% polypeptides.Otherwise the yield of rough polypeptides was 31.41% ± 0.47%,which contained 84.03% ± 0.85%polypeptides,0.11% ± 0.01% glycosaminoglycans and 5.68% ± 0.15% total-sugar.In one word,the co-production process for polysaccharides and polypeptides from Ostrea rivularis was feasible,basically the same as the predicted value.
引文
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[19]杨丽芝,杨贤庆,黄卉,等.响应面法优化南海鸢乌贼墨汁多糖提取工艺[J].食品工业科技,2016,37(18):292-296.
[20]陈康,戴志远,沈清,等.响应面法提取南极磷虾磷脂及其分子种组成的测定[J].南方水产科学,2017,13(3):104-112.
[21]孙建瑞,符丹丹,赵君峰,等.响应面法优化Chlamydomonas sp.212胞内多糖发酵工艺及其抑菌活性研究[J].食品科学,2017,38(18):1-7.
[22]鲁伟,任国谱,宋俊梅.蛋白水解液中多肽含量的测定方法[J].食品科学,2005,26(7):169-171.
[23]杨文博,张英华.蛋白质水解度的测定方法研究[J].中国调味品,2014,39(3):88-90.
[24]王剑锋,孙爽,李瑶,等.甲醛滴定法测定牛血清白蛋白的酶解度[J].大连民族学院学报,2012,14(5):441-444.
[25]何钊.白蜡虫多糖分离纯化与抗氧化、免疫活性研究[D].北京:中国林业科学研究院,2014:21-27.
[26]刘义,钱和.1,9-二甲基亚甲基蓝光度法测定海参糖胺聚糖[J].分析试验室,2005,24(6):59-61.
[27]Farndale R W,Buttle D J,Barrett A J.Improved quantitation and discrimination of sulphatedglycosaminoglycans by use of dimethylmethyleneblue[J]. BiochimB iophysA cta,1986,883(2):173-177.
[28]周小双,王锦旭,杨贤庆,等.响应面法优化合浦珠母贝糖胺聚糖提取工艺[J].食品与发酵工业,2016,42(1):238-243.
[2]Wang J,Hu J,Cui J,et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats[J].Food Chemistry,2008,111(2):302-308.
[3]陈慧斌,王梅英,王莉.褶牡蛎成分组成及营养学评价研究[J].宁德师专学报:自然科学版,2009,21(4):342-345.
[4]张显良.2017中国渔业统计年鉴[M].北京:中国农业出版社,2017:23.
[5]郝记明,章超桦,郭顺堂.酶解制备马氏珠母贝肉ACEIP的工艺研究[J].食品科学,2007,28(5):128-131.
[6]Kim S,Wijesekara I.Development and biological activities of marine-derived bioactive peptides:A review[J]. Journal of Functional Foods,2010,2(1):1-9.
[7]Senevirathne M,Kim S. Development of bioactive peptides from fish proteins and their health promoting ability[J].Advances in Food and Nutrition Research,2012,65:235-248.
[8]Zhao Y,Chen P. Natural products with health benefits from marine biological resources[J].Current Organic Chemistry,2014,18(7):777-792.
[9]吴静,胡晓,杨贤庆,等.鸢乌贼酶解产物的抗氧化稳定性与功能特性[J].南方水产科学,2016,12(5):105-111.
[10]牛瑞,崔金香.海洋生物酶解多肽的研究进展[J].化工科技市场,2010,33(3):29-30.
[11]Wang J,Hu J,Cui J,et al.Purification and identification of a ACE inhibitory peptide from oyster proteins hydrolysate and the antihypertensive effect of hydrolysate in spontaneously hypertensive rats[J].Food Chemistry,2008,111(2):302-308.
[12]Miyajima T,Ogawa H,Koike I. Alkali-extractable polysaccharides in marine sediments:Abundance,molecular size distribution,and monosaccharide composition[M]. 2001:1455-1466.
[13]Mirzaei M,Mirdamadi S,Ehsani M R,et al.Purification and identification of antioxidant and ACE-inhibitory peptide from Saccharomyces cerevisiae protein hydrolysate[J]. Journal of Functional Foods,2015,19:259-268.
[14]Je J,Lee K,Lee M H,et al.Antioxidant and antihypertensive protein hydrolysates produced from tuna liver by enzymatic hydrolysis[J]. Food Research International,2009,42(9):1266-1272.
[15]张菊华,杨荣文,刘伟,等.枳实中橙皮苷与辛弗林的工业化联产工艺研究[J].食品与机械,2016,32(11):169-173.
[16]Chen X,Li H,Sun S,et al.Co-production of oligosaccharides and fermentable sugar from wheat straw by hydrothermal pretreatment combined with alkaline ethanol extraction[J].Industrial Crops and Products,2018,111:78-85.
[17]牛改改,游刚,张晨晓,等.响应面法优化近江牡蛎多糖提取工艺[J].南方农业学报,2016,47(6):994-1000.
[18]郑亚旭,石璇,李欣遥,等.超滤膜分离牡蛎多糖工艺研究[J].安徽农业科学,2016,44(19):104-106.
[19]杨丽芝,杨贤庆,黄卉,等.响应面法优化南海鸢乌贼墨汁多糖提取工艺[J].食品工业科技,2016,37(18):292-296.
[20]陈康,戴志远,沈清,等.响应面法提取南极磷虾磷脂及其分子种组成的测定[J].南方水产科学,2017,13(3):104-112.
[21]孙建瑞,符丹丹,赵君峰,等.响应面法优化Chlamydomonas sp.212胞内多糖发酵工艺及其抑菌活性研究[J].食品科学,2017,38(18):1-7.
[22]鲁伟,任国谱,宋俊梅.蛋白水解液中多肽含量的测定方法[J].食品科学,2005,26(7):169-171.
[23]杨文博,张英华.蛋白质水解度的测定方法研究[J].中国调味品,2014,39(3):88-90.
[24]王剑锋,孙爽,李瑶,等.甲醛滴定法测定牛血清白蛋白的酶解度[J].大连民族学院学报,2012,14(5):441-444.
[25]何钊.白蜡虫多糖分离纯化与抗氧化、免疫活性研究[D].北京:中国林业科学研究院,2014:21-27.
[26]刘义,钱和.1,9-二甲基亚甲基蓝光度法测定海参糖胺聚糖[J].分析试验室,2005,24(6):59-61.
[27]Farndale R W,Buttle D J,Barrett A J.Improved quantitation and discrimination of sulphatedglycosaminoglycans by use of dimethylmethyleneblue[J]. BiochimB iophysA cta,1986,883(2):173-177.
[28]周小双,王锦旭,杨贤庆,等.响应面法优化合浦珠母贝糖胺聚糖提取工艺[J].食品与发酵工业,2016,42(1):238-243.
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