Optimization of Fermentation Process for Human-like Collagen Production of Recombinant Escherichia coli Using Response Surface Methodology

In order to improve the production of human-like collagen III(HLC III)by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation process parameters.Three variables(induction time,inoculum age and pH),which have significant effects on HLC III production,were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design,a relationship between HLC III production and three significant factors was obtained,and the optimum levels of the three variables were as follows:induction time 3.2h,inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An average 9.68 g·L1HLC III production was attained in the validation experiment under optimized condition,which was 80%higher than the yield of 5.36 g·L1before optimization.

Optimization of Fermentation Process for Hyaluronic Acid Production by Streptococcus zooepidemicus

Hyaluronic acid （HA） is a high molecular weight glycosaminoglycan consisting of alternating D-glucuronic acid and N-acetylglueasamine and plays ex- tremely important roles in many biological processes. In this study, we optimized fermentation process for the production of HA by Streptococcus zooepidemicus ATCC35246, including fermentation broth composition and various fermentation parameters. The experimental results showed that the optimal fermentation broth composition was： glucose 45 g/L, yeast extract 10 g/L, tryptone 12 g/L, KH2PO4 2 g/L, K2HPO4 . 3H20 2 g/L, MgSO4 · 7H2O 2 g/L, and （NH4 ）2SO4 0.4 g/L. The optimal parameters involved in fermentation was： liquid volume 20%, pH 6. 0, rotation speed 180 r/min, fermentation temperature 35 ℃, fermentation duration 18 h, CTAB concentration 25 mg/L. Under the optimized conditions, the yield of HA was 0. 305 g/L, which was dramatically improved by 43.87% compared to that of 0. 212 g/L before optimization.

Recent Progress of Commercially Available Biosensors in China and Their Applications in Fermentation Processes

Biosensors, which are the products of the biotechnology industry, are among the key projects of the 7th, 8th, and 9th Fiveyear Plans of China Science ＆ Technology Developing Programs, respectively, and they play an important role in developing and reforming traditional biotechnology. SBA series biosensor analyzer, as the only one commercial biosensor in China, has attracted lots of attention in the process of information gathering and measurement for biological industry with the development of technology and society. In this paper, we presented an overview of the most important contributions dealing with the monitoring of the biochemical analytes in fermentation processes using SBA series biosensor analyzers in China. Future trends of the biosensor analyzer in China were also mentioned in the last section.

Conditional Optimization of Laccase Production by Whiterot Fungi through Fermentation

Phanerochaete chrysosporium was selected as the production strain of laccase,and the effects of stirring speed,ventilation volume,culture temperature,inoculation amount and initial p H of medium on laccase production by liquid fermentation in cylinder were studied. On the basis of single factor test,an orthogonal test was carried out to find optimal conditions for laccase production P. chrysosporium through liquid fermentation. These results showed that the stirring speed of fermentation cylinder had the highest effect on laccase production,and the optimal conditions were shown as follows： the temperature at 28 ℃,the rotating speed at 300 r/min,the ventilation volume of 5 L/min（ ventilation ratio of 1.0 vvm）,the initial p H of medium of 5,and the inoculation amount of 15%,which gave the highest laccase level of 14. 86 U/ml.

Method of Soft-Sensor Modeling for Fermentation Process Based on Geometric Support Vector Regression

The soft-sensor modeling for fermentation process based on standard support vector regression(SVR) needs to solve the quadratic programming problem(QPP) which will often lead to large computational burdens, slow convergence rate, low solving efficiency, and etc. In order to overcome these problems, a method of soft-sensor modeling for fermentation process based on geometric SVR is presented. In the method, the problem of solving the SVR soft-sensor model is converted into the problem of finding the nearest points between two convex hulls (CHs) or reduced convex hulls (RCHs) in geometry. Then a geometric algorithm is adopted to generate soft-sensor models of fermentation process efficiently. Furthermore, a swarm energy conservation particle swarm optimization (SEC-PSO) algorithm is proposed to seek the optimal parameters of the augmented training sample sets, the RCH size, and the kernel function which are involved in geometric SVR modeling. The method is applied to the soft-sensor modeling for a penicillin fermentation process. The experimental results show that, compared with the method based on the standard SVR, the proposed method of soft-sensor modeling based on geometric SVR for fermentation process can generate accurate soft-sensor models and has much less amount of computation, faster convergence rate, and higher efficiency.

Whole process reclamation and utilization of wastes produced in the biological fermentation industry

Wastes yielded in the vintage process and the biological fermentation of itaconic acid and sodium gluconate of a winery in Shandong, such as grain stillage, melon lees, cornstarch protein residues, itaconic acid mother liquid, itaconic acid mycelium and sodium gluconate mycelium, were studied. High-activity biological protein feed, foliar fertilizer and irrigation fertilizer were generated from these wastes by applying biological/microbial technologies. Meanwhile, a whole set of technological pathways was put forward. As a result, the optimal economical and social benefits can be obtained with low natural resource consumption and environmental costs by converting wastes into useful matters. In conclusion, through the utilization of limited resources in the whole process of reclamation and utilization of wastes, the harmony promotion can be achieved between the economic system and the natural ecosystem.

OPTIMAL FEED STRATEGY FOR FED-BATCH GLYCEROL FERMENTATION DETERMINED BY MAXIMUM PRINCIPLE

Optimal glucose feed strategy for glycerol fed-batch fermentation was investigated by Pontryagin’s maximum principle to maximize the final glycerol yield. The problem was solved by a nonsingular control approach by selecting the culture volume as the control variable, then the general optimal feed profile was numerically determined.

葡萄干酿制葡萄酒的工艺优化

目的研究并优化用普通葡萄干酿造葡萄酒的工艺。方法通过单因素和正交试验研究料水比、果胶酶添加量、酵母菌接种量和发酵温度对葡萄酒品质的影响,并优化酿造工艺。结果单因素试验结果表明,料水比、果胶酶的添加量以及发酵温度主要影响葡萄酒的色度,对残糖浓度和酒精度的影响不明显。相比之下,酵母菌接种量对这些特性的改变贡献较小。正交试验表明,发酵温度、酵母菌接种量和料水比对酒精度有显著影响,且果胶酶添加量明显影响感官评分。在最佳的发酵条件下[发酵温度20℃、酵母菌接种量0.50 g/L、果胶酶的添加量25 mg/L、料水比1:6(g/mL)],可以酿造出酒精度约为12.3%vol,感官评分约为90.5分的高品质葡萄酒。结论普通葡萄干酿制葡萄酒可行性较高,不同因素组合选择余地大,葡萄酒品质符合国家标准,酿造工艺具有一定的商业价值。

淡豆豉的多菌种协同发酵工艺优化

目的优化淡豆豉多菌种(环状芽孢杆菌、黑曲霉、少根根霉)协同发酵工艺,为淡豆豉发酵工艺改进和创新提供参考。方法以发酵时间、发酵温度、混合菌种比例、接种量为考察因素,以总黄酮、大豆苷元、黄豆黄素、染料木素的含量为考察指标,通过单因素考查对淡豆豉多菌种协同发酵工艺进行优化。结果淡豆豉多菌种协同发酵最佳工艺条件为:发酵温度30℃,发酵时间8 d,接种量5%,混合菌种接种比例为环状芽孢杆菌∶黑曲霉∶少根根霉=1∶2∶1。结论淡豆豉多菌种协同发酵工艺可避免杂菌干扰、利于工艺标准化,可为淡豆豉工艺改进和创新以及为中药多菌种协同发酵工艺研究提供参考。

一株耐高温β-胡萝卜素降解菌的筛选及发酵工艺优化

以提升烟叶香韵为目的,对4株产香酵母菌进行发酵培养,通过产香感官评价、酶活分析、烟叶发酵成分分析(GC-MS法),筛选得到1株β-胡萝卜素降解产香酿酒酵母(Saccharomyces cerevisiae)SC35。方法:①4株酵母菌发酵产香效果感官评价;②CS35菌株β-胡萝卜素降解效果分析、样品萃取及GC-MS靶向测试分析;③酶活测定、酶促反应体系研究;④菌株SC35发酵工艺优化及响应面优化。SC35在70℃高温环境下能有效加速β-胡萝卜素的降解,降解产物为1,3-环己二酮、氧化异佛尔酮、β-紫罗兰酮等,通过单因素试验和响应面法优化菌株SC35的最佳培养基配方中各组分添加量依次为葡萄糖2.01%,NaNO20.20%,K2HPO40.10%,MgSO40.05%,FeSO4·7H2O 0.001%,KCl 0.05%;最佳发酵条件为发酵温度27.65℃,发酵时间72 h,接种量0.99%,摇床转速140 r/min,装液量100 mL/250 mL,pH值5.5(自然)。采用优化后发酵工艺SC35粗酶液降解效率可达86%以上。

干酪乳杆菌LK-1的发酵特性及其在菠萝果汁中的应用

为利用干酪乳杆菌制备发酵菠萝果汁饮品,本研究对干酪乳杆菌LK-1(Lactobacillus casei LK-1,L. casei LK-1)的生长曲线、产酸、耐酸和耐糖等特性进行研究分析,并以活菌数和总酸为指标,通过单因素实验及响应面法考察L. casei LK-1发酵菠萝果汁的最佳条件。结果表明,L. casei LK-1在pH5~7和0%~10%质量分数葡萄糖环境下,具备良好的生长繁殖能力;在酸性(pH3)和高糖(40%质量分数葡萄糖)环境中培养3 h,其存活率分别为76%和71%;在MRS培养基中培养48 h,产酸量为5.35 g/kg,产酸能力良好,适用于果汁的发酵。L. casei LK-1发酵菠萝果汁的最佳条件为:初始pH6.8,发酵温度37℃,接种量1%(v/v),发酵时间30 h,此条件下制备的发酵菠萝果汁饮品活菌数为8.99±0.04 lg CFU/mL,总酸含量为7.16±0.26 g/kg,且色泽鲜亮,酸甜适口,风味较好,为L. casei LK-1在发酵食品中的进一步应用提供了理论和技术依据。

番茄酵素发酵工艺优化及营养成分变化分析

以番茄为原料,植物乳杆菌为发酵菌种,超氧化物歧化酶(SOD)和总酸为评价指标,采用单因素试验和Box-Behnken试验优化番茄酵素的发酵工艺,并分析监测番茄酵素发酵过程中营养成分和相关酶活性变化。结果表明,在最佳接种量0.3%、发酵温度37℃、初始pH值5.5、发酵时间40 h的条件下,番茄酵素SOD活性达147.63 U/g,总酸含量为48.60 g/kg。发酵过程中,pH值和可溶性固形物含量均随着发酵时间的增加而持续下降,总酸含量持续上升,还原糖、总酚含量及SOD活性、脂肪酶活性均先上升后下降,番茄红素含量先持续下降后上升再下降;同时对各营养成分与酶活性进行相关性分析,结果表明各营养成分之间存在一定相关性。

酿酒酵母和异常汉逊酵母混合发酵黄酒工艺研究

以酿酒酵母和异常汉逊酵母为黄酒发酵菌种,分别考察2种酵母对18°Bx糖化液的发酵特性,结果表明酿酒酵母产酒精(9.9%vol)能力强于异常汉逊酵母(6.8%vol),异常汉逊酵母产挥发酯(2.41 g/L)显著高于酿酒酵母(0.53 g/L)。比较混菌发酵途径,顺序混合发酵酒精度(8.1%vol)略低于同时混合发酵酒精度(8.7%vol),挥发酯含量(1.59 g/L)显著高于同时混合发酵(0.56 g/L)(P<0.05),结合发酵过程理化指标和感官评价结果,选择顺序混合发酵途径较为合适。通过单因素和Box-Benhnken中心组合试验设计,选择主发酵温度、pH值、酵母接种量3个因素对顺序混合发酵黄酒产品的挥发酯含量进行响应面优化,结果显示:顺序混合发酵黄酒的最佳参数为:发酵温度31.5℃,接种量1.0×10~7个/mL,pH 4.2。验证试验表明,在此条件下顺序混合发酵黄酒的挥发酯含量为(1.66±0.028)g/L,与预测值1.70 g/L相差不大,比酿酒酵母单独发酵提高213%。本研究为非酿酒酵母在黄酒发酵上的应用奠定了基础。

甜瓜酸奶发酵工艺优化及其品质分析

该研究以纯牛奶和甜瓜汁为原料制备甜瓜酸奶,以酸度、持水力和感官评分为考察指标,在单因素试验的基础上采用响应面法对甜瓜酸奶的发酵工艺进行优化,并对其贮藏特性、抗氧化活性、流变特性、微观结构以及挥发性风味成分进行分析。结果表明,甜瓜酸奶的最佳发酵工艺条件为:甜瓜汁添加量7%,赤藓糖醇添加量7%,发酵时间8 h,接种量0.3%。在此优化条件下,甜瓜酸奶酸度为82.62°T,持水力为81.13%,感官评分为90.70分。在贮藏21 d后,甜瓜酸奶的感官评分、酸度、持水力分别为73.0分、88.4°T、68.2%;甜瓜酸奶具有均一、连续、紧密蛋白质网络结构和更好的流变性能;共检出33种挥发性风味成分,包括7种醇类、9种酯类、7种醛类、4种酮类、2种酸类和4种其他类化合物,相对含量分别为19.64%、28.43%、27.70%、11.69%、9.18%、3.36%。甜瓜汁的添加能改善酸奶的品质,赋予酸奶甜瓜香气。

多菌种混合发酵神农架猕猴桃果醋的工艺优化及风味分析

以神农架猕猴桃为原料,通过多菌种混合发酵制备猕猴桃果醋。通过单因素及正交试验优化其酒精、醋酸发酵工艺条件,采用氨基酸分析仪及气质联用(GC-MS)技术分别分析果醋的氨基酸、挥发性风味物质组成。结果表明,最佳酒精发酵工艺为:酵母菌接种量0.9%,乳酸菌接种量7%,初始可溶性固形物19°Bx,发酵温度27℃。在此优化条件下,酒精度为(8.82±0.57)%vol,乳酸含量为(12.15±0.73)mg/mL。最佳醋酸发酵工艺为:发酵时间8 d,醋酸菌接种量12%,初始酒精度7%vol,发酵温度32℃。在此优化条件下,醋酸含量为(4.34±0.17)g/100 mL,感官评分为80分。多菌种混合发酵猕猴桃果醋共检出15种氨基酸,包含7种必需氨基酸;共检出37种挥发性风味物质,包括酯类17种,酸类5种,醛类2种,酮类5种,苯类2种,醇类4种,酚类1种,其他类1种。综上,多菌种混合发酵有利于促进氨基酸和挥发性风味物质的形成,从而提升果醋的品质。

鱼类肠道益生地衣芽孢杆菌FA6发酵工艺的响应面法优化

研究以地衣芽孢杆菌FA6活菌数为指标,采用单因素实验和响应面法实验优化了地衣芽孢杆菌FA6发酵培养基和发酵条件,获得了地衣芽孢杆菌FA6最佳发酵培养基:70 g/L可溶性淀粉、40 g/L酪蛋白胨、2 g/L K2HPO4、1 g/L KH_(2)PO_(4)、2 g/L Na_(2)HPO_(4)、2 g/L MgSO_(4)和8 g/L NaCl;以及最佳发酵条件:初始pH 7.0、接种量1%、发酵温度28.8℃、发酵时间31h、装液量50 mL/250 mL和转速160 r/min。在此发酵培养基和发酵条件下,地衣芽孢杆菌FA6活菌数为7.97×10^(9)CFU/mL,较优化前提高了8.1倍。最佳发酵工艺的获得,将能够为鱼源芽孢杆菌的高密度生产,产业化应用提供基础。

大肠杆菌发酵低廉生物质产乙醇酸及其分离纯化研究

碳中和背景下全生物基乙醇酸得到了广泛的关注,但其高昂发酵成本、匮乏的下游分离纯化工艺研究,限制了后续的工业生产应用。该研究以重组大肠杆菌为对象,采用统计学方法在摇瓶水平上对发酵培养基成分进行优化,并基于5 L发酵罐进行初步放大验证和下游分离纯化研究。最终利用低廉的玉米芯水解液等成分,摇瓶水平乙醇酸产量达到4.77 g/L,较初始培养基提高2.58倍;5 L罐乙醇酸产量达42 g/L,提高至优化前的1.4倍;通过初步的生物分离纯化,成功获得生物基乙醇酸,晶体纯度达到99.3%,符合市售标准要求。该研究通过发酵培养基优化、5 L发酵罐放大验证以及下游生物分离纯化,初步构建了生物基乙醇酸的低成本生产体系,为生物基乙醇酸的工业化生产应用奠定基础。

高产抗菌脂肽芽孢杆菌的分离鉴定筛选及发酵工艺优化

为了筛选出高产抗菌脂肽的芽孢杆菌并确定其最佳发酵工艺条件,研究从传统发酵豆酱中分离和鉴定产抗菌脂肽芽孢杆菌菌株,通过正交试验研究了发酵接种量、发酵填液量、发酵时间、发酵温度等发酵条件对芽孢杆菌脂肽产量的影响。结果表明,从9份东北传统发酵豆酱中分离筛选得到27株芽孢杆菌,经16S rDNA鉴定,其中6株芽孢杆菌具有合成脂肽的基因sfp、fenB和ituA。通过单位菌体脂肽产量和抑菌效果测定发现,贝莱斯芽孢杆菌SN-20和解淀粉芽孢杆菌SN-46表现优异,单位菌体脂肽产量达106和72 mg,且对革兰氏阳性和阴性指示菌均有较强的抑菌效果。正交试验发现贝莱斯芽孢杆菌SN-20最佳发酵工艺为:接种量3%,发酵填液量20%,发酵时间36 h,发酵温度32℃;枯草芽孢杆菌SN-46最佳发酵工艺为:接种量2%,发酵填液量40%,发酵时间24 h,发酵温度32℃。在此条件下发酵的两株芽孢杆菌单位菌体脂肽产量优化前为106.11和76.23 mg/g,优化后分别提高了21.85%和23.84%。研究结果有效地增加了芽孢杆菌抗菌脂肽的产量。

植物酸奶品质优化的研究进展

植物酸奶的出现为牛乳蛋白过敏、高胆固醇、关注动物福利和环境保护人群带来了福音。然而,植物酸奶因存在乳清析出、异味及酸度不高等品质问题降低了消费者的喜爱度,因此对植物酸奶品质优化进行研究在拓宽植物基食品的商业应用方面意义重大。本文概述了不同来源植物酸奶营养、风味及质构上存在的主要品质问题,着重阐述了植物酸奶品质调控的相关手段及措施,包括配方改良、加工条件优化、发酵方式改善三方面的作用方式及效果,探讨了植物酸奶体系稳定、植物蛋白溶解性提高、凝胶网络交联加强等方面的机理。不同调控措施对植物酸奶品质的改良效果存在差异、各有优缺点,有时需多种方法配合使用,以最大化改善植物酸奶品质问题。最后展望了一些潜在的植物酸奶品质调控措施,以期为生产高品质植物酸奶提供指导方向。

响应面法优化荞麦食醋酒精发酵工艺

为了提高荞麦食醋发酵过程中酒精发酵阶段的乙醇产量,以发酵醪酒精度为响应值,以固液比、酵母接种量、接种温度、发酵温度、发酵时间等因素为自变量,利用单因素结合响应面试验,优化荞麦醋酒精发酵工艺条件。结果表明,最佳酒精发酵工艺为固液比1∶4(g∶mL)、酵母接种量30%、酵母接种温度30℃,发酵温度28℃,发酵时间7 d。在此优化条件下,酒精度为9.52%vol,比优化前(8.51%vol)提高了11.9%。