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黑曲霉丝氨酸羧肽酶的重组表达及其水解大豆蛋白的效果分析 被引量:1
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作者 喻豆 潘力 王斌 《现代食品科技》 CAS 北大核心 2022年第1期173-180,158,共9页
丝氨酸羧肽酶能够剪切肽链C末端的氨基酸,在大豆蛋白脱苦中有着广泛的应用。该研究将黑曲霉中3个丝氨酸羧肽酶基因(CPG、CPF、CPA)在低背景的无孢黑曲霉HL-1中进行重组表达。利用启动子PnaⅡ、终止子Ttef和营养缺陷型筛选标记pyr G构建... 丝氨酸羧肽酶能够剪切肽链C末端的氨基酸,在大豆蛋白脱苦中有着广泛的应用。该研究将黑曲霉中3个丝氨酸羧肽酶基因(CPG、CPF、CPA)在低背景的无孢黑曲霉HL-1中进行重组表达。利用启动子PnaⅡ、终止子Ttef和营养缺陷型筛选标记pyr G构建分别含自身信号肽和糖化酶信号肽的丝氨酸羧肽酶表达载体;利用PEG介导的转化法转化无孢黑曲霉宿主HL-1,构建了丝氨酸羧肽酶重组表达菌株。通过摇瓶发酵筛选得到高表达重组菌株HL-CPG,其丝氨酸羧肽酶酶活达到163.71 U/mL。利用6ⅹHis标签进行镍柱亲和层析得到单一的丝氨酸羧肽酶CPG并研究了其酶学性质,该酶最适反应温度为40℃,最适p H为3.5,Cu^(2+)具有明显抑制效果。另外,将重组表达的丝氨酸羧肽酶CPG与胃蛋白酶复配水解大豆蛋白,水解液中疏水性氨基酸Leu、Tyr和Phe的含量分别增加606.47μg/m L、434.06μg/m L和205.11μg/m L。综上所述,该研究在黑曲霉中成功实现丝氨酸羧肽酶的高效表达,对大豆蛋白水解液脱苦处理工艺具有一定的借鉴意义。 展开更多
关键词 丝氨酸羧肽酶 黑曲霉 重组表达 蛋白脱苦
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Enzymatic Hydrolysis of Defatted Mackerel Protein with Low Bitter Taste 被引量:4
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作者 HOU Hu LI Bafang ZHAO Xue 《Journal of Ocean University of China》 SCIE CAS 2011年第1期85-92,共8页
Ultrasound-assisted solvent extraction was confirmed as a novel,effective method for separating lipid from mackerel pro-tein,resulting in a degreasing rate (DR) of 95% and a nitrogen recovery (NR) of 88.6%.To obtain p... Ultrasound-assisted solvent extraction was confirmed as a novel,effective method for separating lipid from mackerel pro-tein,resulting in a degreasing rate (DR) of 95% and a nitrogen recovery (NR) of 88.6%.To obtain protein hydrolysates with high ni-trogen recovery and low bitter taste,enzymatic hydrolysis was performed using eight commercially available proteases.It turned out that the optimum enzyme was the ‘Mixed enzymes for animal proteolysis’.An enzyme dosage of 4%,a temperature of 50℃,and a hydrolysis time of 300 min were found to be the optimum conditions to obtain high NR (84.28%) and degree of hydrolysis (DH,16.18%) by orthogonal experiments.Glutamic acid was the most abundant amino acid of MDP (defatted mackerel protein) and MDPH (defatted mackerel protein hydrolysates).Compared with the FAO/WHO reference protein,the essential amino acid chemical scores (CS) were greater than 1.0 (1.0 1.7) in MDPH,which is reflective of high nutritional value.This,coupled with the light color and slight fishy odor,indicates that MDPH would potentially have a wide range of applications such as nutritional additives,functional ingredients,and so on. 展开更多
关键词 MACKEREL HYDROLYSIS ULTRASOUND defatted BITTER amino acid composition
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