粗酶水相提取大豆油过程中蛋白酶解与油脂释放状态
|
摘要
为了考察粗酶水相提取大豆油过程中蛋白酶解和油脂释放状态,监测酶解过程中总油、总蛋白提取率以及蛋白相对分子质量变化,同时采用扫描电镜、考马斯亮兰与苏丹红Ⅲ染色显微观察技术研究酶解过程中油脂和蛋白在豆粉、水解液、乳状液中的分布状态。结果表明:随酶解时间延长,蛋白酶解程度加深,油脂释放量提升,水解蛋白中相对分子质量大于33 000 Da的多肽含量逐渐降低,而相对分子质量小于5 500 Da的短肽含量增加。酶解6 h时,总蛋白和总油提取率均达到最高,分别为90. 1%和94. 2%,此时蛋白水解液中短肽(<5 500 Da)含量最高,占49. 63%,相对分子质量大于33 000 Da的多肽仅占21. 15%。扫描电镜图显示由于酶解作用豆粉内的蛋白和油脂逐渐被分解和释放,原本充盈的细胞转变为空的残渣。光镜图显示,伴随豆粉酶解,水解液中蛋白含量逐渐增加,释放的油脂在水解液和乳状液中均有分布,随酶解时间延长油滴数量增加、粒径增大。
In order to investigate the status of protein hydrolysis and oil release during crude enzyme-assisted aqueous extraction of soybean oil,the extraction rates of total oil and total protein were monitored,and relative molecular weight of the protein was detected by gel chromatography. Meanwhile,scanning electron microscopy( SEM),light microscopy with Coomassie brilliant blue and Sudan red Ⅲ staining were used to reveal the distribution of oil and protein in soybean powder,hydrolysate and emulsion during the enzymolysis. The results showed that with the increase of enzymolysis time,the degree of protein hydrolysis deepened,and the oil release amount increased. The content of polypeptides with relative molecular weight higher than 33 000 Da gradually decreased,while the content of short polypeptides with relative molecular weight less than 5 500 Da increased. The highest extraction rates of total oil and total protein were obtained at enzymolysis time 6 h,which were 94. 2% and 90. 1% respectively,the content of short peptides( < 5 500 Da) in protein hydrolysate was the highest,accounting for 49. 63%,and the content of polypeptides with relative molecular weight higher than 33 000 Da was only 21. 15%.SEM indicated that the protein and oil in soybean powder were gradually degraded and released by enzymolysis so that the original intact cells turned into empty residues. Light microscope pictures showed that with the hydrolysis of soybean powder,the protein content in hydrolysate increased progressively,and the oil released was distributed in hydrolysate and emulsion. The number and diameter of oil droplets increased with the extension of enzymolysis time.
In order to investigate the status of protein hydrolysis and oil release during crude enzyme-assisted aqueous extraction of soybean oil,the extraction rates of total oil and total protein were monitored,and relative molecular weight of the protein was detected by gel chromatography. Meanwhile,scanning electron microscopy( SEM),light microscopy with Coomassie brilliant blue and Sudan red Ⅲ staining were used to reveal the distribution of oil and protein in soybean powder,hydrolysate and emulsion during the enzymolysis. The results showed that with the increase of enzymolysis time,the degree of protein hydrolysis deepened,and the oil release amount increased. The content of polypeptides with relative molecular weight higher than 33 000 Da gradually decreased,while the content of short polypeptides with relative molecular weight less than 5 500 Da increased. The highest extraction rates of total oil and total protein were obtained at enzymolysis time 6 h,which were 94. 2% and 90. 1% respectively,the content of short peptides( < 5 500 Da) in protein hydrolysate was the highest,accounting for 49. 63%,and the content of polypeptides with relative molecular weight higher than 33 000 Da was only 21. 15%.SEM indicated that the protein and oil in soybean powder were gradually degraded and released by enzymolysis so that the original intact cells turned into empty residues. Light microscope pictures showed that with the hydrolysis of soybean powder,the protein content in hydrolysate increased progressively,and the oil released was distributed in hydrolysate and emulsion. The number and diameter of oil droplets increased with the extension of enzymolysis time.
引文
[1]吴海波,江连洲,程建军,等.粗酶水解全脂豆粉提取油脂和蛋白[J].农业工程学报,2011,27(4):376-382.
[2]吴海波,齐宝坤,江连洲,等.挤压膨化及粗酶添加对水相制取大豆油和蛋白质的影响[J].中国食品学报,2017,17(10):71-79.
[3]张毛赛,楼飞,张瑞娟,等.频率、温度和大豆分离蛋白对南极磷虾虾肉糜介电特性的影响[J].食品与发酵工业,2017,43(8):212-218.
[4]沈金荣,史梦珂,邓泽元,等.大豆复合植物蛋白饮料配方优化及其理化性质[J].食品工业科技,2018,39(2):175-181.
[5]王才立,张效伟,张钊.乳化型大豆分离蛋白对冰淇淋质构的影响及工艺优化[J].食品与发酵工业,2017,43(8):219-223.
[6]ZHANG H,BARTLEY G E,ZHANG H,et al.Peptides identified in soybean protein increase plasma cholesterol in mice on hypercholesterolemic diets[J].J Agric Food Chem,2013,61(35):8389-8395.
[7]MOURA J M L N D,CAMPBELL K,MAHFUZ A,et al.Enzyme-assisted aqueous extraction of oil and protein from soybeans and cream de-emulsification[J].J Am Oil Chem Soc,2008,85(10):985-995.
[8]ROSENTHAL A,PYLE D L,NIRANJAN K,et al.Combined effect of operational variables and enzyme activity on aqueous enzymatic extraction of oil and protein from soybean[J].Enzyme Microb Tech,2001,28(6):499-509.
[9]LI P F,GASMALLA M A A,LIU J,et al.Characterization and demusification of cream emulsion from aqueous extraction of peanut[J].J Food Eng,2016,185:62-71.
[10]HANMOUNGJAI P,PYLE D L,NIRANJAN K.Enzymeassisted water extraction of oil and protein from rice bran[J].J Chem Technol Biot,2002,77(7):771-776.
[11]DE MOURA J M L N,DE ALMEID N M,JUNG S,et al.Flaking as a pretreatment for enzyme-assisted aqueous extraction processing of soybeans[J].J Am Oil Chem Soc,2010,87(12):1507-1515.
[12]TOWA L T,KAPCHIE V N,WANG G,et al.Quantity and quality of free oil recovered from enzymatically disrupted soybean oleosomes[J].J Am Oil Chem Soc,2011,88(10):1581-1591.
[13]韩宗元,江连洲,李杨,等.水酶法提取大豆油的扩大试验研究[J].中国粮油学报,2015,30(2):37-42.
[14]HARDOUIN K,BURLOT A S,UMAMI A,et al.Biochemical and antiviral activities of enzymatic hydrolysates from different invasive French seaweeds[J].J Appl Phycol,2014,26(2):1029-1042.
[15]李杨,江连洲,许晶,等.挤压膨化预处理水酶法提取大豆油工艺的研究[J].中国油脂,2009,34(4):6-10.
[16]LAMSAL B P,JOHNSON L A.Separating oil from aqueous extraction fractions of soybean[J].J Am Oil Chem Soc,2007,84(8):785-792.
[17]LAMSAL B P,REITMEIER C,MURPHY P A,et al.Enzymatic hydrolysis of extruded-expelled soy flour and resulting functional properties[J].J Am Oil Chem Soc,2006,83(8):731-737.
[18]CZUBINSKI J,SIGER A,LAMPART-SZCZAPA E.Digestion susceptibility of seed globulins isolated from different lupin species[J].Eur Food Res Technol,2016,242(3):391-403.
[19]JMIN D M B,MAURER D,YAO L,et al.Characteristics of oil and skim in enzyme-assisted aqueous extraction of soybeans[J].J Am Oil Chem Soc,2013,90(7):1079-1088.
[20]KEN S,HIDEKAZU I,CHIE K,et al.Peptide-based tumor inhibitor encoding mitochondrial p14(ARF)is highly efficacious to diverse tumors[J].Cancer Sci,2016,107(9):1290-1301.
[21]ORTIZ-MARTINEZ M,OTERO-PAPPATHEODOROUJ T,SERNA-SALDVER S O,et al.Antioxidant activity and characterization of protein fractions and hydrolysates from normal and quality protein maize kernels[J].JCereal Sci,2017,76:85-91.
[22]陈星星,胡晓,李来好,等.抗疲劳肽的研究进展[J].食品工业科技,2015,36(4):365-369.
[23]ZHANG S,LIU X,LU Q,et al.Enzymatic demulsification of the oil-rich emulsion obtained by aqueous extraction of peanut seeds[J].J Am Oil Chem Soc,2013,90(8):1261-1270.
[2]吴海波,齐宝坤,江连洲,等.挤压膨化及粗酶添加对水相制取大豆油和蛋白质的影响[J].中国食品学报,2017,17(10):71-79.
[3]张毛赛,楼飞,张瑞娟,等.频率、温度和大豆分离蛋白对南极磷虾虾肉糜介电特性的影响[J].食品与发酵工业,2017,43(8):212-218.
[4]沈金荣,史梦珂,邓泽元,等.大豆复合植物蛋白饮料配方优化及其理化性质[J].食品工业科技,2018,39(2):175-181.
[5]王才立,张效伟,张钊.乳化型大豆分离蛋白对冰淇淋质构的影响及工艺优化[J].食品与发酵工业,2017,43(8):219-223.
[6]ZHANG H,BARTLEY G E,ZHANG H,et al.Peptides identified in soybean protein increase plasma cholesterol in mice on hypercholesterolemic diets[J].J Agric Food Chem,2013,61(35):8389-8395.
[7]MOURA J M L N D,CAMPBELL K,MAHFUZ A,et al.Enzyme-assisted aqueous extraction of oil and protein from soybeans and cream de-emulsification[J].J Am Oil Chem Soc,2008,85(10):985-995.
[8]ROSENTHAL A,PYLE D L,NIRANJAN K,et al.Combined effect of operational variables and enzyme activity on aqueous enzymatic extraction of oil and protein from soybean[J].Enzyme Microb Tech,2001,28(6):499-509.
[9]LI P F,GASMALLA M A A,LIU J,et al.Characterization and demusification of cream emulsion from aqueous extraction of peanut[J].J Food Eng,2016,185:62-71.
[10]HANMOUNGJAI P,PYLE D L,NIRANJAN K.Enzymeassisted water extraction of oil and protein from rice bran[J].J Chem Technol Biot,2002,77(7):771-776.
[11]DE MOURA J M L N,DE ALMEID N M,JUNG S,et al.Flaking as a pretreatment for enzyme-assisted aqueous extraction processing of soybeans[J].J Am Oil Chem Soc,2010,87(12):1507-1515.
[12]TOWA L T,KAPCHIE V N,WANG G,et al.Quantity and quality of free oil recovered from enzymatically disrupted soybean oleosomes[J].J Am Oil Chem Soc,2011,88(10):1581-1591.
[13]韩宗元,江连洲,李杨,等.水酶法提取大豆油的扩大试验研究[J].中国粮油学报,2015,30(2):37-42.
[14]HARDOUIN K,BURLOT A S,UMAMI A,et al.Biochemical and antiviral activities of enzymatic hydrolysates from different invasive French seaweeds[J].J Appl Phycol,2014,26(2):1029-1042.
[15]李杨,江连洲,许晶,等.挤压膨化预处理水酶法提取大豆油工艺的研究[J].中国油脂,2009,34(4):6-10.
[16]LAMSAL B P,JOHNSON L A.Separating oil from aqueous extraction fractions of soybean[J].J Am Oil Chem Soc,2007,84(8):785-792.
[17]LAMSAL B P,REITMEIER C,MURPHY P A,et al.Enzymatic hydrolysis of extruded-expelled soy flour and resulting functional properties[J].J Am Oil Chem Soc,2006,83(8):731-737.
[18]CZUBINSKI J,SIGER A,LAMPART-SZCZAPA E.Digestion susceptibility of seed globulins isolated from different lupin species[J].Eur Food Res Technol,2016,242(3):391-403.
[19]JMIN D M B,MAURER D,YAO L,et al.Characteristics of oil and skim in enzyme-assisted aqueous extraction of soybeans[J].J Am Oil Chem Soc,2013,90(7):1079-1088.
[20]KEN S,HIDEKAZU I,CHIE K,et al.Peptide-based tumor inhibitor encoding mitochondrial p14(ARF)is highly efficacious to diverse tumors[J].Cancer Sci,2016,107(9):1290-1301.
[21]ORTIZ-MARTINEZ M,OTERO-PAPPATHEODOROUJ T,SERNA-SALDVER S O,et al.Antioxidant activity and characterization of protein fractions and hydrolysates from normal and quality protein maize kernels[J].JCereal Sci,2017,76:85-91.
[22]陈星星,胡晓,李来好,等.抗疲劳肽的研究进展[J].食品工业科技,2015,36(4):365-369.
[23]ZHANG S,LIU X,LU Q,et al.Enzymatic demulsification of the oil-rich emulsion obtained by aqueous extraction of peanut seeds[J].J Am Oil Chem Soc,2013,90(8):1261-1270.