豆豉纤溶酶生产菌液体发酵条件的优化

以DCFM9突变株为出发菌株,对其液体发酵条件进行了优化,得到了最佳发酵条件.优化后发酵培养基组成:木糖为20 g,大豆蛋白胨为20 g,KH2PO4为1g,K2HPO4·3H2O为4 g,Ca-Cl2为0.4 g,MgSO4·7H2O为0.5 g,水为1 000 mL;摇瓶发酵的最佳工艺条件为pH7.4,接种量2%,种龄24 h,装液量50 mL/250 mL,37℃,180 r/min,发酵12 h加入1g/L的Tween 40.在此条件下发酵72 h,产酶量最高可达3011.08 U/mL.     

Effect of fermented rice culture on the microbiological,biochemical and sensory characteristics of low‐salt douchi,a traditional Chinese fermented soybean condiment

A low‐salt (4% NaCl, w/w) douchi supplemented with different concentration of fermented rice culture (FRC) of 0%, 5%, 10%, 15% and 20% (v/w) was produced, and the effect of FRC on microbiology, biochemistry and sensory quantity was investigated. Results indicated that FRC had a significant effect on low‐salt douchi. Compared with the control (added 0% FRC), in the other treatments, the counts of yeast and lactic acid bacteria increased by about 50%, whereas the moulds decreased by about 30%; the pH reduction was close to 4.7; total acid increased to about 1.300 g per 100 g; reducing sugar maintained a decrease to 21.07 mg per g; amino nitrogen increased to about 0.900 g per 100 g, but not above the control (1.013 g per 100 g). In general terms, total free amino acid contents and three organic acids were higher in low‐salt products than four commercial products. Sensory evaluation of products supplemented with 10% and 15% FRC revealed better overall acceptability compared with other treatments and a commercial product.     

Changes of Composition and Angiotensin I-Converting Enzyme-Inhibitory Activity During Douchi Fermentation

Douchi is a fermented soybean food, which is popular in China. Composition and angiotensin I-converting enzyme-inhibitory activity of Douchi fermented with Mucor wutungkiao were studied in this work. The results showed that the nutritional potency of protein, fat, and soluble polysaccharide was improved in Douchi fermentation. The angiotensin I-converting enzyme-inhibitory activity of Douchi that ripening for 2 weeks showed the highest angiotensin I-converting enzyme-inhibitory activity. Then this sample was purified with Sephadex G-25 gel chromatography, and the amino acids and angiotensin I-converting enzyme-inhibitory activity of the purified fractions were studied. The results suggested that hydrophobic amino acids were assumed to cause the main increase of the angiotensin I-converting enzyme-inhibitory activity in Douchi.     

Antioxidant properties of protein hydrolysate from Douchi by membrane ultrafiltration

Protein extracted from Douchi was hydrolyzed by alcalase based on a single-factor experiment and response surface methodology was employed to optimize hydrolysis conditions. The optimum conditions were determined to be: 63℃ extraction temperature, 1.4% quantities of enzyme and substrate, and 1.7 h extraction time. The optimal protein hydrolysate was subsequently subjected to ultrafiltration, and the fraction of molecular weight 10–50 kDa was identified as the most potent contributor to antioxidant activity. Amino acid analysis indicated that acidic amino acids were the most abundant in all fractions of the protein hydrolysate. Infrared spectroscopy showed there were no significant differences in secondary structure between fractions.     

豆豉溶栓酶工程菌的发酵条件及重组溶栓酶的分离纯化

对豆豉溶栓酶工程菌株WB DFE5进行了摇瓶发酵条件的研究 ,确定了发酵的优化条件为 :可溶性淀粉 2 % ,酪蛋白 2 % ,大豆蛋白胨 0 5 % ,酵母粉 0 15 % ,K2 HPO40 2 5 % ,KH2 PO40 0 5 % ,CaCl2 0 0 2 % ,MgSO40 0 5 % ,pH7 0。通过硫酸铵分段盐析、离子交换和凝胶过滤 ,从 1L工程菌株WB DFE5的发酵液中分离纯化出 12 5mg重组豆豉溶栓酶 ,酶的比活为 5 918 7IU/mg。     

Investigation of the microbial changes during koji‐making process of Douchi by culture‐dependent techniques and PCR‐DGGE

Douchi as a traditional fermentation soyfood with an acquired taste had been appreciated by consumers for thousands of years in China and few Asian countries. Unfortunately, few studies had been conducted on the changes of microbial community during the koji‐making process, and whether the pathogenic microorganisms involving the fermentation process is still unclear. Therefore, the PCR‐denaturing gradient gel electrophoresis (DGGE) targeting the 16S and 18S rRNA genes was applied to characterise the dynamic changes of Bacteria, Bacilli and Fungi during koji‐making process. The results of DGGE showed that eleven species of bacteria, nine species of bacilli and seven species of Fungi had been detected during the koji‐making progress, of which the Bacillus subtilis and Aspergillus oryzae belonged to the dominant microorganisms. Also, eleven strains were isolated from Koji‐making samples and were identified as B. subtilis, Bacillus amyloliquefaciens, A. oryzae, Brachybacterium sp., Aspergillus niger, Staphylococcus saprophyticus, Micrococcus sp., Staphylococcus gallinarum, Absidia corymbifera, Saccharomyces cerevisiae, Malassezia sp.; Our results revealed that pathogenic microorganisms were involved in the koji‐making process, but the probiotic microorganisms occupied the dominant position of community in the koji‐making process.     

基于高通量测序分析不同浏阳豆豉中真菌的多样性

建立测定酱油中铅的氢化物原子荧光光谱法,同时以酱油为样本进行精密度和准确度测定,以及对酱油能力验证盲样考核样进行不确定度分析。结果表明,铅含量在0～100.00 ng/mL呈线性关系,相关系数（R）为0.999 8,该方法的检出限为0.74 μg/L,定量限为2.22 μg/L,精密度试验结果相对标准偏差（RSD）为1.16%～4.48%,回收率为93.3%～106.6%。2个盲样考核样不确定度结果分别为（0.940±0.092） mg/L和（0.460±0.035） mg/L,能力验证考核结果为满意。研究结果表明,采用水质分析质量控制工作程序对氢化物原子荧光光谱法测定酱油中铅进行分析质量控制,保证实验结果误差在可控制的范围内,获得高度可信的分析结果。     

永川毛霉型豆豉在发酵过程中微生物总量与区系变化规律

采用巢式聚合酶链式反应配合变性梯度凝胶电泳技术对永川豆豉发酵过程中微生物区系生态演化进行解析。结果表明：永川豆豉在制曲过程中霉菌和细菌呈对数增长,进入后发酵阶段菌落总数快速下降,并保持在较低水平。永川豆豉在制曲前期有多种乳酸菌生长,后期乳酸菌受霉菌增长抑制,种类减少。在后发酵阶段奥德赛芽孢杆菌（Bacillus odysseyi）、乳酸杆菌（Lactobacillus oligofermentans）和乳酸杆菌（Lactobacillus lindneri）是优势菌群。同时在制曲初期也发现了费格森埃希菌（Escherichia fergusonii）等杂菌生长。永川豆豉制曲阶段优势霉菌是总状毛霉（Mucor racemosus）,同时伴有有性根霉（Rhizopus sexualis）、匍枝根霉（Rhizopus stolonifer）、大孢联轭霉（Syzygites megalocarpus）、米根霉（Rhizopus oryzae）的生长,后发酵阶段有接合酵母（Zygosaccharomyces sp.）的参与。     

闽北豆豉生产过程中营养成分的变化

研究了闽北豆豉生产过程中水分、还原糖、蛋白质、氨基酸态氮及赖氨酸等营养成分含量的变化。结果表明,闽北豆豉发酵7 d后,水分含量由53.34%下降到27.34%;还原糖含量的变化呈先增后减趋势,先由1.42%上升至2.50%后又降低至1.82%;蛋白质含量略有上升,从36%上升至41.77%;氨基酸态氮含量由0.45 g/100 g上升至0.90 g/100 g;赖氨酸含量由2.45 mg/100 g上升至19.23 mg/100 g。     

毛霉型豆豉后发酵工艺条件的优化研究

豆豉后发酵是风味物质形成、营养成分变化及功能因子形成的重要阶段,以感官评价和理化指标评分作为考察指标,采用正交试验对毛霉型豆豉后发酵工艺条件进行优化试验,并对最佳条件进行验证试验,结果表明：当食盐质量分数为9%、白酒添加量为5%、发酵温度为45 ℃、发酵时间为40 d时,毛霉型豆豉的感官评分为14分,理化指标评分为8分。豆豉的氨基酸态氮含量为0.69%,总酸为1.95%,蛋白质为39.20%,还原糖含量为3.60%。