Optimized the Process of Enzymes Coupled with Lactobacillus plantarum and Saccharomyces cerevisiae to Ferment Ginkgo Seeds by Response Surface Methodology
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摘要: 本文旨在运用响应面法优化银杏果的酶菌协同发酵工艺，以期得到一种抗氧化活性高的银杏发酵粉。通过单因素实验探讨了料液比、复合酶制剂添加量、糖化酶和α-淀粉酶添加质量比、酶解时间、混合发酵剂添加量、植物乳杆菌和酿酒酵母添加质量比、发酵时间对银杏发酵粉可溶蛋白含量和DPPH自由基清除效果的影响；并以DPPH自由基清除率为指标，通过响应面分析对银杏果酶菌协同发酵条件进行优化。结果表明，在复合酶制剂添加总量0.3 g/100 g，酶解2 h，发酵12 h的基础上，银杏果酶菌协同发酵最佳工艺为：料液比1：6 g/mL，糖化酶和α-淀粉酶添加质量比2：1，混合发酵剂添加总量3.12 g/100 g，植物乳杆菌和酿酒酵母添加质量比1：1，测得DPPH自由基清除率为69.7%±1.1%，与模型预测值基本一致。由最佳工艺制得的银杏发酵粉（GFP）与未处理的银杏粉（G）相比，DPPH自由基清除率增加了257.4%；可溶蛋白含量4.47±0.13 g/100 g，增加了330%；黄酮含量0.15±0.01 mg/g，增加了50%；多酚含量2.49±0.04 mg/g，增加了118%；银杏酸含量2.96±0.32 μg/g，脱除率达85.4%。说明酶菌协同发酵能显著（P<0.05）提高银杏果的抗氧化活性，制得的银杏发酵粉中可溶蛋白、黄酮、多酚等抗氧化成分含量显著（P<0.05）增加、有毒成分银杏酸含量显著（P<0.05）降低，为银杏果的精深加工提供物质基础。Abstract: This study aimed to optimize the process of enzymes coupled with Lactobacillus plantarum and Saccharomyces cerevisiae to ferment ginkgo seeds by response surface methodology,in order to obtain a kind of ginkgo fermentation powder with high antioxidant activity. The effects of solid-liquid ratio,total amount of compound enzyme,mass ratio of glucoamylase and α-amylase,enzymolysis time,total amount of mixed fermentation agent,mass ratio of Lactobacillus plantarum and Saccharomyces cerevisiae,fermentation time on soluble protein content and DPPH radical scavenging capacity of ginkgo fermentation powder(GFP)were studied by single factor experiment,and based on the DPPH radical scavenging capacity,response surface analysis was used to optimize the process conditions of Lactobacillus plantarum and Saccharomyces cerevisiae coupled with enzymes to ferment ginkgo seeds. The results showed that on the basis of adding 0.3 g/100 g of compound enzyme,enzymolysis 2 h and fermentation 12 h,the best technology of cooperation of bacteria and enzyme fermentation was as follows:1:6 of solid-liquid ratio,2:1 of glucoamylase and α amylase,3.12 g/100 g of mixed fermentation agent,1:1 of Lactobacillus plantarum and Saccharomyces cerevisiae,the DPPH radical scavenging capacity was 69.7%±1.1%,which was basically consistent with the model prediction. Compared with the untreated ginkgo powder(G),the DPPH free radical scavenging rate of Ginkgo fermentation powder(GFP)increased by 257.4%,the soluble protein content was 4.47±0.13 g/100 g,increased by 330%,the flavone content was 0.15±0.01 mg/g,increased by 50%,the polyphenol content was 2.49±0.04 mg/g,increased by 118%,the ginkgolic acid content was 2.96±0.32 μg/g,the removal rate was 85.4%. It showed that enzymes coupled with Lactobacillus plantarum and Saccharomyces cerevisiae fermentation could significantly improve the antioxidant activity of ginkgo seeds,and the contents of antioxidant components as soluble protein,flavonoids and polyphenols in the GFP were significantly increased,and the contents of toxic components ginkgolic acid was significantly decreased,which provided the material basis for the deep processing of ginkgo seeds.
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