响应面法优化芽孢杆菌FC96培养基组分的研究

为了提高芽孢杆菌FC96在液体发酵培养基中的生物量,采用响应面法对其培养基组分进行优化。通过单因素试验确定对芽孢杆菌FC96具有最佳增菌效果的碳源、氮源和无机盐,利用响应面分析法优化培养基组分的最佳配比。试验结果表明,单因素试验确定的最佳碳源、氮源和无机盐分别是葡萄糖、牛肉膏和磷酸二氢钾,响应面法优化芽孢杆菌FC96最佳培养基组成为葡萄糖12.11g/L、牛肉膏23.31g/L和磷酸二氢钾2.33g/L。模型预测的最高活菌数为2.85×109cfu/mL。在未优化培养基中的活菌数为2.32×109cfu/mL。在优化的最佳培养基中,验证试验的最高活菌数为2.97×109cfu/mL,菌数比优化前提高了28%,试验值与预测值的误差为4.21%。     

玫瑰花发芽糙米酵素工艺研究

本研究以优质糙米、玫瑰花为主要原料,应用发酵技术,研究一款集玫瑰花与发芽糙米为一体的酵素。以感官评分为指标,通过单因素试验确定玫瑰花的最佳添加量,并对酵母菌接种量、发酵温度和发酵时间通过响应面优化得到最佳工艺参数。利用优化工艺发酵后对产品理化和微生物指标进行测定,参照标准对产品质量进行评估。经实验得到:在玫瑰花与发芽糙米比例为1∶3的培养基中接入4%的酵母菌31℃发酵8.5h后得到的酵素口味最佳,该酵素pH值(4.32)、NaCl(0.07g/100g)、总酸(3.22g/100g)、游离氨基酸(53.21g/100g)、γ-氨基丁酸(749.73mg/kg)、粗多糖(0.27g/100g)和乳酸菌[3×106CFU/g(mL)]符合限量规定,大肠杆菌、金黄色葡萄球菌、沙门氏菌、霉菌均未检出,产品香甜、软糯、安全。本研究为玫瑰花的利用及新型发芽糙米的酵素的研制提供参考。     

益生酪酸菌的固态培养

以活菌数为考察指标,研究酪酸菌在3种不同固态基质(豆粕、麸皮、玉米粉)中的生长情况。结果显示：豆粕作为基质时酪酸菌生长最佳,最高活菌数可达47×106CFU/g,麸皮次之,达39×106CFU/g,玉米粉最差,仅为34×106CFU/g。在此基础上,采用响应面法对酪酸菌固态发酵的培养基及发酵工艺进行优化。首先通过Plackett-Burman试验对酪酸菌培养基成分进行筛选,从10个因素中筛选到了4个主要影响因素,即硫酸铵、麦芽糖、硫酸镁和含水量。然后采用Box-Behnken试验设计得出硫酸铵、麦芽糖、硫酸镁的添加量分别为1.5%、0.8%、0.02%,含水量为55%时培养效果最佳,酪酸菌活菌数最高可达到约76×106CFU/g。同样采用Box-Behnken试验设计对影响酪酸菌固体发酵的工艺条件进行优化,当发酵时间为24h、温度30℃、接种量14mL/100g时,酪酸菌培养效果最佳,活菌数达到约11×107CFU/g。     

一株清酒乳杆菌的分离、鉴定及培养基优化

从传统食品谷物发酵液中筛选出1株乳杆菌,通过生理生化试验与菌落和菌株形态观察以及16S rRNA测序鉴定为清酒乳杆菌。以MRS培养基为基础培养基,活菌数为指标,改变基础培养基组成的成分及添加量,在此基础上,通过响应面优化培养基组成。结果表明最优培养基组成为鱼蛋白胨14.50 g/L、酵母浸粉6.50 g/L、葡萄糖28.00 g/L、柠檬酸三铵1.50 g/L、磷酸二氢钾2.00 g/L、硫酸镁0.20 g/L、硫酸锰0.12 g/L、吐温-80 1.20 mL/L,此条件下,活菌数达到3.125×10⁹ CFU/mL,相比MRS发酵培养基活菌数(1.980×10⁸ CFU/mL)提高了1 478.28%。该优化培养基具有活菌数高和经济方便的特点,为清酒乳杆菌的工业化生产提供借鉴。     

应用响应面优化泡菜用微生物的富集培养基

采用植物乳杆菌与肠膜明串珠菌为目的菌株,以活菌数为指标,通过选用不同的碳源、氮源、生长因子进行单因素实验得出最优项,再进行响应面分析得出最优的组合。结果表明,由方程可以得出蔗糖取1.04g/100mL,酵母粉含量取0.41g/100mL,麦芽汁添加量为4.58mL/100mL时,肠膜明串珠菌活菌数可达到10.04×109cfu/mL;麦芽浸粉含量为1.61g/100mL,水解大豆蛋白胨1.47g/100mL,番茄汁7.36mL/100mL时,植物乳杆菌活菌数可达到最大值30.13×109cfu/mL。     

动物双歧杆菌乳亚种HCS04-002发酵条件优化

为实现动物双歧杆菌乳亚种（Bifidobacterium animalis subsp. lactis）HCS04-002的高活菌数培养,获得其生长的最适发酵条件,对发酵工艺和发酵培养基分别进行优化。以菌泥收率为考察指标,通过单因素及正交试验对培养温度、接种量和初始pH值等发酵工艺参数进行了优化;以发酵液活菌数为响应值,通过单因素试验和Box-Behnken试验优化发酵培养基。结果表明,动物双歧杆菌乳亚种HCS04-002最佳发酵条件为：培养温度39 ℃、接种量3%、初始pH值为7.2;最佳发酵培养基组分为：酵母蛋白胨24 g/L、酵母浸出物30 g/L、葡萄糖19 g/L、乳糖11 g/L。在此优化条件下,菌株HCS04-002菌悬液活菌数达2.73×109 CFU/mL。     

响应面法优化长双歧杆菌增殖培养基

为了提高长双歧杆菌发酵液中的活菌数,对其增殖培养基进行响应面优化。通过单因素试验筛选出长双歧杆菌的最佳碳源为乳糖,并发现低聚木糖、菊糖、低聚异麦芽糖、低聚果糖、苯丙氨酸、蛋氨酸、脯氨酸、谷氨酸及赖氨酸均能显著促进长双歧杆菌的生长。利用Design Expert 8.06软件设计Plackett-Burman 试验筛选出影响长双歧杆菌生长的3个最重要因子,通过Box-Behnken试验及响应面分析确定3个因子的最佳添加量为：低聚木糖1.7g/L、菊糖3.6g/L、脯氨酸0.4g/L,用优化后的增殖培养基培养长双歧杆菌,18h后其活菌数达(1.75±0.02)×109CFU/mL,比优化前提高了95.64%。     

枯草芽孢杆菌产蛋白酶发酵培养基的优化

在摇瓶发酵条件下,利用响应面法优化枯草芽孢杆菌BS3菌株产蛋白酶的培养基,在单因素实验的基础上,采用Plackett-Burman实验设计(PB)对影响枯草芽孢杆菌BS3产蛋白酶的培养基组分进行了筛选,确定主要影响因子为玉米浆、氯化铵和玉米淀粉.然后通过最陡爬坡实验、中心组合实验设计和响应面分析法,确定了主要的影响因子及其浓度.优化后的培养基组成为:玉米淀粉1.586g/100g,玉米浆3.170/100g,氯化铵1.998/100g,其他组分维持低添加量,在此培养基条件下,活菌数为12.640×109cfu/g,蛋白酶活为162.309U/g.     

鼠李糖乳杆菌LP216高密度发酵培养基优化

为提高鼠李糖乳酸杆菌LP216生产效益,通过单因素试验、Plackett-Burman（PB）试验和Box-Behnken（BB）试验,对菌株LP216发酵培养基进行优化。结果表明,最佳培养基配方为酵母提取物16 g/L、蛋白胨13 g/L、葡萄糖30 g/L、牛肉膏5 g/L、CH3COONa 4 g/L、吐温80 1.5 g/L、MgSO4·7H2O 0.25 g/L、柠檬酸二胺2.0 g/L、K2HPO4 2.0 g/L、MnSO4 0.2 g/L。在此优化条件下,菌株LP216活菌数达8.9×109 CFU/mL,是对照组（MRS培养基）活菌数（3.28×109 CFU/mL）的2.71倍。     

响应面法优化萌发藜麦芽乳发酵工艺

以萌发藜麦芽为原料,研究发酵条件对藜麦芽发酵乳酸度和活菌数的影响。选用植物乳杆菌和干酪乳杆菌2种混合菌进行发酵,通过单因素试验、响应面优化试验探究菌种比例、接种量和发酵时间对发酵的影响。结果表明,萌发藜麦芽乳混合菌发酵最佳工艺条件为植物乳杆菌和干酪乳杆菌比例2.5∶1、接种量3%、发酵时间10.3 h,得到的萌发藜麦芽发酵乳酸度为85.32°T,活菌数为9.21(lg(CFU/m L)),与预测值吻合,表明萌发藜麦芽的匀浆发酵培养基适合乳酸菌生长。     

醋酸菌麸曲制备工艺的优化

以巴氏醋杆菌C9-4(Acetobacter pasteurianum strain C9-4)为麸曲发酵菌株,应用响应面法优化了醋酸菌麸曲的制备工艺条件。结果表明,其最佳制备条件为:乙酸添加量1.03 mL/100 g、乙醇添加量2.09 mL/100 g、接种量4.09mL/100g,该条件下醋酸菌菌落总数达到1.38×10~8 CFU/g。应用该条件,在食醋企业对醋酸菌麸曲进行逐级扩大培养,扩培过程中麸曲醋酸菌总数均能达到10~8 CFU/g以上。     

基于MTT比色和响应面法优化侧孢短芽孢杆菌最大活菌数培养条件

目的：探索获得侧孢短芽孢杆菌（Brevibacillus laterosporus）最大活菌数的最佳培养基成分及培养条件。方法：在建立MTT比色法与平板计数法的相关回归方程基础上,对获得最大活菌数的侧孢短芽孢杆菌最适培养基成分（碳源、氮源、无机盐）和培养条件（初始pH、温度、接种量、磷酸二氢钾）进行优化。结果：MTT比色法与平板计数法对活菌数测定结果表现出显著的线性关联（R²>0.999）;麦芽糖、氯化钙、初始pH、磷酸二氢钾为显著影响因子,最佳发酵条件为麦芽糖8.75 g/L,氯化钙0.17 g/L,初始pH 7.07、磷酸二氢钾3.73 g/L,此条件下活菌数为8.12×10⁸ CFU/mL,与理论活菌数（8.25×10⁸ CFU/mL）无显著差异。结论：基于MTT比色和响应面法优化侧孢短芽孢杆菌最大活菌数培养条件,优化后的活菌数较优化前提高了3.02倍。     

发酵糙米的加工工艺

对用酵母发酵糙米的工艺进行研究。以糙米粉为试验材料,添加水、蜂蜜、盐、大麦芽粉为底物,采用活性酵母液进行发酵试验。以淀粉酶活力作为试验指标,设计包含培养基加水量、发酵温度、时间和接种量四个因素水平,通过正交试验确定糙米发酵培养基的最佳配方和最佳发酵条件。优化的工艺条件为加水量为150%,酵母接种量为4%,发酵温度为30℃,发酵时间为为4 h。糙米酵素中的α-淀粉酶含量最高,达到485 U/g。     

Optimization of Calvatia gigantea mycelia production from distillery wastewater

The focus of wastewater management has evolved from treatment technology into resource recovery, which enables one to minimize contaminants and to generate value‐added products. Calvatia gigantea is used not only as a source of food, but has also been used as a traditional Chinese medicine for thousands of years. In this study, the mycelial production of C. gigantea was studied under submerged fermentation conditions using non‐pretreated distilled wastewater from Chinese liquor production. The fermentation medium composition was optimized using response surface methodology involving a Box–Behnken design. Fermentation conditions were optimized using an orthogonal experimental design. The optimized medium composition was the non‐pretreated distilled wastewater of Chinese liquor supplemented with cornflour at 2.35 g/100 mL, (NH₄)₂SO₄ at 1.11 g/100 mL and CuSO₄ at 0.12 g/100 mL. The optimized fermentation conditions were a rotation speed of 150 rpm, an inoculum size of 10% (v/v), a fermentation temperature of 26 °C and a fermentation time 4.5 days. A maximum mycelial biomass yield of 2.75 g/100 mL was achieved using the optimized medium under the optimized conditions. Results from this study suggest that this is a feasible technology for the mycelial production of C. gigantea using the non‐pretreated distilled wastewater from Chinese liquor production. Copyright © 2015 The Institute of Brewing & Distilling     

乳酸菌组合发酵菌种配方及其增殖培养基的优化

为了将甘肃牧区优良乳酸菌株用于直投式酸奶发酵剂的生产,以从甘肃牧区分离所得的乳酸乳球菌乳脂亚种(HM 598685)、嗜酸乳杆菌(HM 598684)、干酪乳杆菌(HM 598683),按照1:0:0、0:1:0、0:0:1、1:1:0、1:2:0、2:1:0、1:0:1、1:0:2、2:0:1、1:1:1、1:2:1、2:1:1、1:1:2的比例进行混合发酵,通过所制酸奶的感官性状和生化特性确定组合发酵的菌种配方,并对混合发酵的增殖培养基进行优化。结果发现,当3株菌比例为1:1:1时,其混合发酵的凝乳时间最短,为8.5h;所得酸奶的活菌数最大,高达7.90×10~8CFU/mL;而其感官评价中滋味气味及组织形态均显著优于其他组合。根据美蓝实验,当基础培养基中使用胰蛋白酶水解乳蛋白时,培养所得乳酸菌的活性最高,且活菌数可达2.93×10~8CFU/mL。通过L_9(3~4)正交设计,确定最佳增殖因子为酵母粉1%、果糖0.5%、吐温0.2%、乙酸钠0.5%,此时培养所得混合菌活菌数达2.11×10~9CFU/mL;各增殖因子对活菌数影响的主次顺序为:酵母粉>乙酸钠>果糖>吐温。     

Development of rice as potential carriers for probiotic Lactobacillus amylovorus

An experiment using a standard strain of Lactobacillus amylovorus TISTR1110 to test for the capability of carbohydrate utilisation of nine cultivars of rice found that L. amylovorus TISTR1110 could utilise carbohydrate from all cultivars and also exhibit nonhaemolytic properties. After testing the viability of the strain under simulated gastrointestinal tract conditions by enumeration after 5 h, the results showed that the amount of L. amylovorus TISTR1110 present was 4.0 × 10⁵ CFU/mL or a 73.78% survival rate. From the evaluation of Thai rice cultivars as carriers in probiotic products, the highest amount of probiotic bacteria was found in baked and encapsulated glutinous Luem Pua Thai sticky rice up to 4.9 × 10⁶ CFU/g or a survival rate of 69.39 ± 0.06%. The second highest rate (4.8 × 10⁵ CFU/g) was found in heat–moisture steamed rice of Riceberry rice cultivar or a survival rate of 59.74 ± 0.11%. After testing the capability of adherence using SEM, the results showed that glutinous Luem Pua sticky rice resulted in considerably high survival rate of probiotic bacteria compared with other rice. Additionally, the microbiological safety test reported that the amounts of contaminants were acceptable.     

Enhancement of Biological Properties of Soymilk by Fermentation

Soymilk has attracted much interest in the food industry because of its health- promoting properties. Fermentation with lactic acid bacteria is known to provide value addition to soymilk by reducing the beany flavor and content of indigestible oligosaccharides and by enhancing the bioavailability of isoflavones, resulting in a nutritious probiotic food product. In the present study, Soymilk fermentation was studied using Lactobacillus acidophilus strain isolated from ragi, and effect of soymilk supplementation on the fermentation characteristics was also investigated. Viable count of 1.99 × 10⁹ CFU/mL sample and 630.1 U/g of β-glucosidase activity were determined in soymilk after 12 h of fermentation. Soymilk supplementation with skimmed milk powder at 5% level gave best results with viable counts of 22.9 × 10⁹ CFU/ml sample or 212.9 × 10⁹ CFU/g solids; 764 U/g of β-glucosidase activity, 1.44% la titratable acidity and pH of 3.9. Daidzein and genistein profile in fermented soymilk showed that 45.8% and 57.5% of respective isoflavones existing as aglycone form, indicating enhancement of the biological property of soymilk through improvement of health-relevant bioactive forms.     

植物乳杆菌ZU018增殖培养基的优化

本研究以植物乳杆菌ZU018为研究对象,最终活菌数为主要参考指标,对其发酵培养基成分进行了优化。采用单因素实验选择碳源、氮源的种类及优化浓度,通过部分因子试验设计初步确定了麦芽糖、酵母浸粉及柠檬酸三铵为培养基中最主要的三个影响因素,进一步运用最陡爬坡试验及Box-Benhnken试验设计对培养基组分进行优化。结果表明,最佳优化培养基配方为:麦芽糖30.03 g/L、酵母浸粉37.50 g/L、牛肉浸粉25.00 g/L、柠檬酸三铵4.39 g/L、磷酸氢二钾2.00 g/L、乙酸钠5.00 g/L、硫酸镁 0.20 g/L、硫酸锰0.05 g/L以及1.00 g/L的吐温80。最终发酵液中植物乳杆菌ZU018活菌数相比优化前提高4.86倍,达到10.67×109 CFU/mL,为后续植物乳杆菌高密度发酵及应用提供了理论依据。     

Factors Affecting Zymomonas mobilis subsp. francensis Growth and Acetaldehyde Production

An experimental plan was designed to determine the incidence of factors encountered during cider production on Zymomonas mobilis subsp. francensis growth and acetaldehyde production. Different factor combinations of pH (3.50 to 4.10), SO₂ addition (0, 50, 100 and 200 mg/L), nitrogen source concentrations (0.5 and 5.0 g/L), polyphenol cider marc extract supplementation (0.25 and 1.00 g/L), temperature (12°C, 18.5°C and 25°C) and inoculation level (10² and 10⁵ CFU/mL) were tested over a 30‐day period at regular time intervals in synthetic medium with ethanol. Viable cell counts and acetaldehyde production were correlated. Individually, and in decreasing significance, the following factors influenced acetaldehyde production: nitrogen source, SO₂ addition, inoculation level, temperature and pH. On the other hand, presence of polyphenol cider marc extract was not significant. A model was determined based on factor interactions. At high contamination levels (10⁵ CFU/mL), conditions leading to a high risk for spoilage were observed at pH values ranging from 3.75 to 4.10, at 0 to 50 mg/L total SO₂ and in the presence of 5 g/L nitrogen source (yeast extract) while temperature did not in fact appear to play a key role. At lower contamination levels (10² CFU/mL), the risk was drastically reduced. The only conditions leading to increased acetaldehyde levels were a high pH 4.1, no added SO₂ and high nitrogen source concentration (5 g/L yeast extract).