产朊假丝酵母发酵油茶果壳水解液生产木糖醇

目的:本文以油茶果壳为原料,利用产朊假丝酵母发酵油茶果壳木糖水解液制备木糖醇,得到产朊假丝酵母发酵油茶果壳木糖水解液制备木糖醇的最佳工艺条件,为油茶果壳综合开发提供理论依据及数据参考。方法:采用稀硫酸水解油茶果壳半纤维素,利用Ca(OH)2和活性炭对水解液进行脱色脱毒,再利用产朊假丝酵母发酵水解液制备木糖醇,在单因素实验的基础上,通过响应面法设计实验优化油茶果壳木糖水解液制备木糖醇的工艺条件。结果:最佳优化结果为:初始p H值5.68,发酵温度28℃,装液量53.88 mL,接种量为10%,木糖醇得率优化值为(16.56±0.25)%。结论:方差分析结果表明,所选模型具有显著性(p0.05),相关系数R2=0.9523,验证试验木糖醇得率为(16.56±0.25)%,与预测值16.81%的误差仅为1.48%,证明所得模型可信,优化工艺参数切实可行。     

紫外诱变季也蒙毕赤酵母原生质体筛选木糖醇高产菌株

目的通过对季也蒙毕赤酵母原生质体进行紫外诱变,筛选遗传性稳定的木糖醇高产菌株。方法制备季也蒙毕赤酵母DQ11的原生质体,计算原生质体生成率及再生率,并对制备过程中的酶浓度(0.5%、1.0%、1.5%和2.0%)、酶解温度(26、28、30、32和34℃)及酶解时间(1、2、3、4和5 h)进行优化。对采用最佳酶解条件制备的原生质体进行紫外诱变,分别检测诱变30、60、90、120和150 s时原生质体的致死率,经重复筛选,采用高效液相色谱法测定菌株发酵液的木糖醇含量,并检测筛选出的木糖醇高产菌株的遗传稳定性。结果最佳酶解条件为:酶浓度1.5%,酶解温度30℃,酶解时间3 h,该条件下制备的原生质体经紫外诱变90 s(最佳诱变时间)后,获得1株高产木糖醇诱变菌株YZ-57,其发酵液的木糖醇含量为9.78 g/L,比出发菌株DQ11提高了186%,且具有良好的遗传稳定性。结论季也蒙毕赤酵母原生质体经紫外诱变,成功获得一株季也蒙毕赤酵母木糖醇高产菌株YZ-57,且具有良好的遗传稳定性。     

茶籽壳酸水解活性炭脱毒发酵生产木糖醇工艺研究

在利用茶籽壳中木质纤维素稀酸水解液发酵生产木糖醇的过程中,水解液须经过脱毒处理,以除去其中抑制菌体生长的有毒物质,本研究采用活性炭对茶籽壳稀酸水解液进行吸附脱毒试验,研究了不同因素对活性炭脱毒的影响,结果表明:在初始p H=2.0,以3%的活性炭添加量,在温度温度为60℃下,脱毒90 min,脱毒率可达67.29%。茶籽壳酸水解液经过石灰中和和活性炭脱毒后,采用酵母菌株直接发酵生产木糖醇。并对发酵条件进行了优化,优化结果为:初始p H为6、接种量为14%、转速200 r/min在温度为30℃下发酵120小时,在该条件下,验证实验以茶籽壳水解液发酵木糖醇产量可达到25.18 g/L。     

高产油脂酵母的筛选及发酵条件的研究

采用苏丹黑染色法对6种酵母菌进行初筛及摇瓶发酵复筛,选出了油脂产量高的假丝酵母。并对假丝酵母摇瓶发酵条件进行优化,确定了最适培养条件为:初始pH值6.0,转速180 r.min-1,接种量10%,发酵周期4 d;最佳碳源为葡萄糖,最佳氮源为酵母膏。假丝酵母在优化条件下发酵后细胞量为16.4 g.L-1、油脂含量为12%、油脂产量为1.97 g.L-1,分别是优化前的142%、171%、245%。     

响应面法优化苦荞麦复合醋的发酵工艺

以产自山西朔州的苦荞麦和运城酥梨作为实验材料，采用液态发酵法制备苦荞复合醋，对醋酸菌的接种量、初始pH、发酵的温度以及摇床转速等4个因素进行单因素试验，以总酸含量作为评价指标。在单因素试验的基础上，利用响应面法优化得到苦荞复合醋醋酸发酵的最佳工艺条件：醋酸菌接种量0.84%、pH 5.12、发酵温度32.62℃，在此条件下发酵获得的苦荞复合醋的总酸含量为2.83 g/100 mL。     

毕赤酵母产重组木聚糖酶发酵条件的优化及其酶学性质

目的优化毕赤酵母工程菌GS115/xyn11A产重组木聚糖酶的发酵条件,并检测其酶学性质。方法采用单因素试验和L(934)正交试验考察摇瓶发酵条件下培养基起始pH值、诱导剂甲醇添加量、诱导温度及诱导时间对产酶活性的影响;并分析重组木聚糖酶的酶学性质。结果影响重组毕赤酵母产酶的因素重要性依次为:培养基起始pH值>诱导时间>诱导温度>甲醇添加量,重组酵母产酶最佳条件为:起始pH值7.5,甲醇添加量1.5%,32℃诱导96 h,在此条件下进行诱导表达重组木聚糖酶的酶活性可达228.35 IU/ml;酶的最适反应温度为50℃,最适反应pH值为5.5,在低于40℃和pH 4.5～7.5的范围内较稳定。结论优化了毕赤酵母产重组木聚糖酶的发酵条件,为木聚糖酶的工业化生产及应用提供了依据。     

表达重组谷氨酸脱羧酶工程菌发酵条件的优化

目的对表达谷氨酸脱羧酶(Glutamate decarboxylase,EC4.1.1.15,简称GAD或GDC)的重组大肠杆菌的发酵条件进行优化。方法通过单因素试验优化表达GAD的重组大肠杆菌B3C1的诱导剂IPTG添加时间、IPTG浓度、诱导时间、诱导温度,再经响应面分析法优化工程菌的发酵条件(氯化钠、胰化蛋白胨、酵母粉、摇床转速、初始pH值、培养时间、接种量、装液量)。结果工程菌的最佳诱导条件为:IPTG添加时间为工程菌接种后4 h,IPTG浓度为0.8 mmol/L,诱导时间为4 h,诱导温度为35℃;工程菌的最佳发酵条件为:酵母粉1.01%,氯化钠0.6%,胰化蛋白胨1.5%,培养时间11.25 h,摇床转速250 r/min,初始pH值6.5,接种量3.0%,装液量11.70 ml。工程菌在该条件下发酵培养,GAD酶活达1 227.33 U,比优化前提高了12.29%。结论已成功对表达GAD的重组大肠杆菌的发酵条件进行了优化,为其工业化生产奠定了基础。     

黄酒糟发酵制备细菌纤维素的技术研究

以黄酒糟酶解液为主要原料,葡糖醋杆菌BC19-2发酵制备细菌纤维素.分别考察了温度、pH、接菌量、酒糟的酶解时间等因子对黄酒糟发酵生产细菌纤维素的影响,可视化表征细菌纤维素制备的微观过程.结果表明,单因素比较优化的最佳发酵条件是酒糟酶解3小时、初始pH6.0、接种量为6％时,在30℃恒温发酵7天,细菌纤维素的产量即达7...     

竹黄水解及发酵制备木糖醇的研究(摘要)

以竹黄为原材料,利用热带假丝酵母发酵竹黄半纤维素水解液生产木糖醇,对半纤维素水解液的制备工艺以及水解液的脱毒处理、菌种的驯化改良、木糖醇发酵过程的优化等关键技术进行了研究。     

乙醇发酵菌种的选育及发酵条件的初步探索试验

对5种燃料乙醇发酵菌种进行了选择,对较好菌种做了耐高温驯化及发酵条件的优化。结果表明,在以葡萄糖为单一碳源的发酵中,相对于休哈塔假丝酵母、嗜单宁管囊酵母、间型脉胞菌、好食脉孢菌,酿酒酵母种液制备时间最短(对数生长期在6～13 h),发酵制取乙醇得率最高;对酿酒酵母驯化后,得到一株可在42℃正常生长的菌株;经研究,该耐高温菌株的较优发酵条件为温度30℃、转速150 r/min、初始pH值5.0、接种量10%。该耐高温驯化后的酿酒酵母不能直接用于水解液发酵制乙醇,还需要经过驯化或菌种改良后方能使用。     

Effect of glucose:xylose ratio on xylose reductase and xylitol dehydrogenase activities from Candida guilliermondii in sugarcane bagasse hydrolysate

The influence of glucose on xylose reductase (XR) and xylitol dehydrogenase (XDH) enzyme activity was evaluated from sugarcane bagasse hydrolysate fermentations with different glucose:xylose ratios (1:25, 1:12, 1:5 and 1:2.5) by employing an inoculum of Candida guilliermondii grown in media containing glucose, a mixture of glucose and xylose, or only xylose as carbon sources. According to the results, the glucose:xylose ratio affected positively this bioconversion and a correlation was not observed between the favourable conditions for xylitol production and the XR and XDH activities. Also, the results were influenced not only by the glucose:xylose ratio in the fermentation medium, but also by the carbon source employed in the growth medium of the inoculum. The optimum condition for xylitol production by C. guilliermondii in sugarcane bagasse hemicellulosic hydrolysate should use hydrolysate with a 1:5 glucose:xylose ratio and inoculum grown in medium containing xylose as the only carbon source. Copyright © 2006 Society of Chemical Industry     

Effect of pH and activated charcoal adsorption on hemicellulosic hydrolysate detoxification for xylitol production

Biotechnological conversion of xylose into xylitol using hydrolysates obtained from the hemicellulosic fraction of lignocellulosic materials is compromised by the presence of compounds released or formed during the hydrolysis process, some of them being toxic to microorganisms. In order to improve the bioconversion of these hydrolysates it is necessary to find methods to reduce their toxicity. In the present work, rice straw hemicellulosic hydrolysate was treated by six different procedures (all of them involving pH adjustment, with or without activated charcoal adsorption), before being used as a fermentation medium for xylitol production. The most effective method of treatment was to increase the initial pH (0.4) to 2.0 using solid NaOH, followed by the addition of activated charcoal (25 g kg^?1) and increase in the pH to 6.5 using solid NaOH. Lignin degradation products were the most inhibitory compounds present in the hydrolysate; their removal was selective and strongly dependent on the pH employed in the treatment. The highest yield of xylitol was 0.72 g g^?1 xylose, with a productivity of 0.55 g dm^?3 h^?1. Copyright © 2004 Society of Chemical Industry     

玉米秸秆半纤维素制备木糖醇的研究

首先采用无污染的碱性过氧化氢法研究了半纤维素的分离与提取,然后对提取的半纤维素分别进行化学水解和酶水解比较,最后研究了水解液发酵制备木糖醇。结果表明,半纤维素分离提取的优化参数为:2%过氧化氢,2%氢氧化钠,加热时间4 h,反应温度75℃。使用CF3COOH水解半纤维素所得木糖含量为67%~73%,水解率为76%~84%,稀盐酸预处理半纤维素再化学水解所得木糖含量高达88%,水解率上升至大约90%。半纤维素的酶水解实验表明,木聚糖酶的水解专一性高于半纤维素酶,木聚糖酶水解率为38%~60%。在水解液发酵实验中,酶水解液的木糖醇转化率高于化学水解液。另外,通过浓缩半纤维素水解液,提高发酵液的木糖初始浓度,有利于菌株生长,可以提高木糖醇转化率。研究对于玉米秸秆半纤维素制备化学品具有一定的指导意义。     

麦草酸水解及水解液木糖发酵的研究

采用甲酸/盐酸水解体系水解麦草,发现 65℃ 水解 0.5 h 可实现麦草中半纤维素的充分水解。选用热带假丝酵母发酵麦草甲酸水解液制取木糖醇。分别研究了在不同浓度甲酸及甲酸根条件下D-木糖的发酵效果,发现在 2 g/L 的甲酸及 5 g/L 的甲酸根条件下,D-木糖实现发酵并得到最大产率的木糖醇,高浓度的甲酸及甲酸根都会抑制D-木糖的发酵。采用D311型阴离子交换树脂脱除甲酸根,实现了麦草水解液的发酵,木糖醇最高得率为 16.88%(木糖醇/木糖)。     

真菌发酵酒精废弃物合成单细胞蛋白基质及菌种筛选

为了优选利用酒精废弃物生产单细胞蛋白（single cell protein,SCP）的最佳基质和菌种组合,从而实现其高值化利用,本文选取了黄酒加工废弃物（黄酒糟和米浆水）以及木薯酒精加工废弃物（木薯原糟和木薯酒糟上清液）4种基质作为发酵底物,并以产朊假丝酵母（Candida utilis）和白地霉（Geotrichum candidum）2种常见菌种作为筛选对象,通过摇瓶发酵实验,考察不同基质对菌种产SCP的影响,同时对比不同菌种利用酒精废弃物发酵合成SCP的能力,从而选出最优发酵基质和菌株。结果表明,以黄酒糟和米浆水按质量分数6%配比,同时接种产朊假丝酵母和白地霉进行混菌发酵4天时,SCP的产率及产量均达最优值,干物质产率可达3.91,产量可达90.22g/L,粗蛋白含量可达53.3%,且发酵产物中的氨基酸成分齐全,作为蛋白饲料具有良好的应用前景。     

ORP调控对马克斯克鲁维酵母发酵纤维素水解液的影响

采用通气的方式进行氧化还原电位（oxidation-reduction potential,ORP）调控,研究在添加抑制物条件下Kluyveromyces marxianus 1727-5利用葡萄糖、木糖以及两者混合体系的发酵性能,在此基础上考察了ORP调控策略对玉米秸秆水解液发酵的影响。研究结果表明：在调控ORP策略下,多种混合抑制物对酵母生长代谢造成的损害得以有效改善,细胞活性高,木糖、葡萄糖代谢速率加快。葡萄糖与木糖共发酵时,ORP调控至-150 mV时,葡萄糖发酵时间缩短近30%,木糖醇浓度由3 g·L^-1增加到10 g·L^-1。ORP调控策略也同样能有效缓解玉米秸秆水解液中较高浓度的多种抑制物对酵母细胞的胁迫,ORP为-100 mV时,相比于对照组,在保持终点乙醇不变的情况下,葡萄糖的发酵时间缩短了22%;木糖消耗由4.88 g·L^-1增至10.27 g·L^-1,木糖醇得率也由0.20 g·g^-1提高至0.48 g·g^-1。     

响应面法优化以重组大肠杆菌发酵生产类人胶原蛋白过程

In order to improve the production of human-like collagen Ⅲ (HLC Ⅲ) by fed-batch culture of recom-binant Escherichia coli BL21, the Plackett-Burman and Box-Behnken design were applied to optimize the fermen-tation 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 aver-age 9.68 g·L~(-1)HLC Ⅲ production was attained in the validation experiment under optimized condition, which was 80% higher than the yield of 5.36 g·L~(-1) before optimization.     

Xylitol production by Candida tropicalis from corn cob hemicellulose hydrolysate in a two‐stage fed‐batch fermentation process

BACKGROUND: Xylitol, a sugar alcohol widely used in food and pharmaceutical industries, can be produced through biological reduction of xylose present in hemicellulose hydrolysates by Candida tropicalis. However, the aeration rate and by‐products originating from hemicellulose hydrolysis strongly inhibit the production of xylitol in a fermentation process. A two‐stage fed‐batch fermentation system was developed to reduce these inhibitory effects and to improve xylitol production from corn cob hemicellulose hydrolysates by C. tropicalis. RESULTS: Results of batch fermentations indicated that high xylitol production could be obtained from C. tropicalis at an initial xylose concentration of 80 g L^?1 in corn cob hydrolysate medium at an aeration rate of 0.4 vvm at the micro‐aeration stage. In the two‐stage fed‐batch fermentation process, 96.5 g L^?1 xylitol was obtained after 120 h, giving a yield of 0.83 g g^?1 and a productivity of 1.01 g L^?1 h^?1, which were 12.16% and 65.57% higher than those in a batch fermentation. CONCLUSION: High xylitol production can be achieved in a two‐stage fed‐batch fermentation process, in which the negative effects of aeration rate and inhibitory compounds on xylitol formation can be considerably reduced. Copyright © 2011 Society of Chemical Industry     

海洋微生物溶菌酶的发酵优化与中试生产

以海洋细菌S-12-86为试验菌株,采用摇瓶发酵优化的方式,研究培养基组分(碳源、氮源、碳源与氮源的比例、金属离子)与发酵条件(培养温度、接种体积分数、装液体积分数、起始pH值、产酶周期)对海洋微生物溶菌酶产量的影响,并进行中试放大试验。结果表明:该菌产酶最佳培养基组分为:葡萄糖10 g/L,蛋白胨5 g/L,MgSO45 g/L,CaCl22 g/L;最适发酵培养温度为30℃,接种体积分数为4.0%,装液体积分数为10.0%,起始pH值为8.0,发酵周期24 h。海洋细菌S-12-86发酵优化后的产酶量(25636.8 U/mL)较优化前的产酶量(14454.4 U/mL)提高了75.4%。海洋微生物溶菌酶中试发酵的产酶量达26697.87 U/mL。说明摇瓶发酵优化条件可以应用于海洋微生物溶菌酶中试生产上。     

Clarification of a wheat straw‐derived medium with ion‐exchange resins for xylitol crystallization

BACKGROUND: Xylitol bioproduction from lignocellulosic residues comprises hydrolysis of the hemicellulose, detoxification of the hydrolysate, bioconversion of the xylose, and recovery of xylitol from the fermented hydrolysate. There are relatively few reports on xylitol recovery from fermented media. In the present study, ion‐exchange resins were used to clarify a fermented wheat straw hemicellulosic hydrolysate, which was then vacuum‐concentrated and submitted to cooling in the presence of ethanol for xylitol crystallization. RESULTS: Sequential adsorption into two anion‐exchange resins (A‐860S and A‐500PS) promoted considerable reductions in the content of soluble by‐products (up to 97.5%) and in medium coloration (99.5%). Vacuum concentration led to a dark‐colored viscous solution that inhibited xylitol crystallization. This inhibition could be overcome by mixing the concentrated medium with a commercial xylitol solution. Such a strategy led to xylitol crystals with up to 95.9% purity. The crystallization yield (43.5%) was close to that observed when using commercial xylitol solution (51.4%). CONCLUSION: The experimental data demonstrate the feasibility of using ion‐exchange resins followed by cooling in the presence of ethanol as a strategy to promote the fast recovery and purification of xylitol from hemicellulose‐derived fermentation media. Copyright © 2008 Society of Chemical Industry