热带假丝酵母发酵法生产木糖醇的研究

目的利用热带假丝酵母研究发酵木糖生产木糖醇的发酵条件。方法采用摇瓶发酵对发酵生产条件,如培养基中初始木糖浓度、接种量和通气量、氮源、pH等进行优化,通过测定发酵液中木糖的残留量、木糖醇的转化率来确定适合的发酵工艺。结果通过实验得到最佳培养基条件为:初始木糖50g/L,蛋白胨5g/L,酵母粉10g/L,硫酸镁0.5g/L、磷酸二氢钾5g/L,硫酸铵1g/L;最佳发酵条件为:pH 6.0,摇瓶发酵装液量50mL/250mL,转速200 r/min,发酵温度30℃,发酵时间28h。结论优化了木糖醇的发酵工艺。     

土壤中产木糖醇酵母菌株的筛选其发酵条件优化

本文以木糖为唯一碳源从土壤中筛选得到可以耐受高浓度木糖的菌株,再经过复筛选出一株高产木糖醇的酵母菌株Y-9。经高效液相色谱(HPLC)和红外扫描分析,确定菌株Y-9发酵利用木糖转化得到的主要产物为木糖醇。通过单因素实验、正交试验等手段,对菌株Y-9发酵产木糖醇的培养基组分和发酵条件进行了优化,进一步提高了目的菌株的木糖醇产率和转化率,确定了菌株Y-9摇瓶发酵木糖转化木糖醇的最优培养基和发酵条件。在木糖初始浓度为200 g/L,氮源为酵母膏3.0 g/L,硫酸铵2.0 g/L,玉米浆10.0 mL/L,硫酸镁0.1 g/L,初始pH为6.0,转速为180 r/min,接种量为4%的条件下,菌株Y-9的木糖醇产率为160 g/L左右,木糖醇生成速率为1.67 g/L.h,木糖/木糖醇转化率达到80%以上,是一株具有良好工业化研究开发价值的木糖醇生产菌株。     

热带假丝酵母发酵生产木糖醇的研究

对热带假丝酵母 (C tropicalis )As2 1 776发酵木糖醇的营养条件进行了初步研究。初始木糖浓度在 80g/L附近时木糖醇转化率较高 ,限制性供氧条件下有利于木糖醇积累。酵母膏和蛋白胨是比较适合产木糖醇的有机氮源 ,而酵母膏更利于酵母细胞生长。培养基中添加 2 g/L的(NH4 ) 2 HPO4 、2～ 6g/L的NaCl、1～ 3g/L的KH2 PO4 、0 1～ 0 3 g/L的MgSO4 ·7H2 O能提高木糖醇的转化率     

驯化和胶囊包裹的假丝酵母LF04发酵玉米芯水解液生产木糖醇(英文)

玉米芯的酸水解液是木糖醇生产的重要原料,但是该水解液中含有糠醛、酚类等对后续微生物发酵有毒害作用的化合物。本研究从土壤中分离了一株似假丝酵母LF01,通过驯化和微胶囊包裹来提高其对水解液的抗性。结果表明通过多次驯化并进行包裹的假丝酵母LF04能在玉米芯水解液中不经任何脱毒处理发酵木糖生产木糖醇。在pH5.5 溶氧为 0.15vvm 的条件下发酵 88h,木糖转化率为 76%,木糖醇浓度达 61.768g/L。远高于其出发菌株。该结果表明采用该方法有望用于木糖醇的工业化生产。     

热带假丝酵母发酵甘蔗渣水解液生产木糖醇

用超声辅助双碱(NaOH/氨水)的方法预处理酶解蔗渣,利用热带假丝酵母发酵蔗渣水解液生产木糖醇,优化发酵的工艺参数。以木糖醇质量浓度为参照指标,分别考察了种龄、接种量、木糖初始质量浓度对热带假丝发酵蔗渣水解液生产木糖醇的影响,得出种龄为26 h,接种量为10%,木糖初始质量浓度为100 g/L为发酵木糖醇最适条件。在最适条件下,热带假丝酵母菌以蔗渣酶解液为原料发酵54 h,最终木糖醇质量浓度达到62.98 g/L。结果证明,热带假丝酵母菌可以有效发酵蔗渣水解液,有一定的研究意义。     

热带假丝酵母(Candida tropicalis)HDY-02发酵玉米芯半纤维素水解液产木糖醇的研究

通过发酵罐批次发酵实验,对C.tropicalis HDY-02发酵木糖合成木糖醇的几个重要发酵参数进行了考察。结果显示,发酵最适温度为35℃,最适发酵pH6.0,最适初始木糖浓度为80 g/L,氧传递系数为KL a 16.5 h-1和18.3 h-1时分别获得最大木糖醇得率为0.73 g/g和最大产率为0.84 g/L·h。因此,为获得更高的木糖醇得率和产率,在发酵过程中采用两阶段的通气策略(0~24 h,KL a为18.3 h-1;24 h之后KL a改为16.5 h-1)。在两阶段通气条件下采用发酵温度35℃,发酵pH6.0,初始木糖浓度80 g/L进行批次发酵,72 h时木糖醇浓度和木糖消耗浓度均达最大值,获得了62.1 g/L木糖醇,0.86 g/L·h产率和0.77 g/g得率。     

一株热带假丝酵母木糖醇高产新菌株的筛选

自木糖厂环境样品中分离出一株编号为1-18的产木糖醇分离物。在初始木糖浓度200g/L的条件下重复循环利用1-18细胞摇瓶发酵,木糖醇转化率高达0.91(木糖醇g/木糖g),接近理论极限值;木糖醇生成速率达6.7g/L·h。无论产物浓度,产物生成速率与产物转化率都达到了令人满意的高水平。1-18发酵液以40ml/kg·d的最大剂量饲喂小白鼠一周,受试动物未出现中毒反应。分离物1-18在木糖醇生产中显然具有重要的生物工程应用潜力。根据常规形态鉴别,生理生化实验测定,以及26S rDNA D1/D2区域核酸序列同源性比较分析,证实分离物1-18属于Candida tropicalis(热带假丝酵母)。分离物1-18已保存于中国典型培养物保藏中心,保藏编号CCTCC M 205067,其26S rDNA D1/D2区域核酸序列在GenBank的核酸序列登记号为DQ176428。     

固定化Amycolatopsis sp.ST2710分段发酵无锡他汀

用海藻酸钠对Amycolatopsis sp.ST2710细胞进行固定化处理。以固定化细胞的机械强度以及分段发酵无锡他汀的转化率为指标,考察了海藻酸钠浓度、CaCl2浓度、固定化时间和包埋菌体量对固定化细胞分段发酵无锡他汀的影响,得到最佳的固定化条件为:海藻酸钠浓度20 g/L、CaCl2浓度10 g/L、固定化时间1 h、包埋菌体量2 mL菌悬液/10 mL凝胶。对固定化细胞分段发酵无锡他汀的特点进行了研究,结果显示:Amycolatopsissp.ST2710经过固定化处理后,大幅度提升了其对底物洛伐他汀的耐受性,显著降低了对发酵培养基中的淀粉需求量,可重复利用。当发酵两阶段pH分别为7.5和5.5、发酵时间分别为48 h和10 h、洛伐他汀添加量3 g/L、转化培养基中淀粉浓度15 g/L时,中间产物Ⅰ对洛伐他汀转化率为64%,无锡他汀对中间产物Ⅰ的转化率为75%。连续发酵5批后,产物转化率仍能维持较高的水平。     

聚乙烯醇质量浓度对复合载体固定化热带假丝酵母颗粒强度、形貌与木糖醇发酵的影响

以聚乙烯醇(PVA)与海藻酸钠这两种载体复合包埋热带假丝酵母细胞,制备成固定化细胞颗粒,研究PVA质量浓度对细胞颗粒的磷酸盐耐受性、机械强度与机械稳定性、形貌及木糖醇发酵性能的影响。磷酸盐耐受性实验结果显示,提高PVA质量浓度能有效改善细胞颗粒的磷酸盐耐受性。对细胞颗粒机械强度的分析表明：发酵前所有的细胞颗粒均有较好的机械强度;但是PVA质量浓度偏低时,发酵后颗粒的机械强度大幅度下降;PVA质量浓度提高到60g/L时,细胞颗粒的机械稳定性较好,发酵后能保持较高的机械强度。扫描电镜分析表明：发酵后的细胞颗粒表面出现破裂或小孔,致使产生游离细胞,但游离细胞的量并非随颗粒机械强度的增加而减少;PVA质量浓度变化对细胞在颗粒中的分布、形态和木糖醇发酵性能影响很大;PVA质量浓度较低时,固定化细胞颗粒外层细胞密布,颗粒中心也有较多细胞,细胞呈饱满的卵圆形;随PVA质量浓度增加,颗粒内部的细胞逐渐减少;PVA质量浓度达到60g/L以上时,细胞主要分布在颗粒外层,且呈现聚集生长的现象,部分细胞发生变形;PVA质量浓度在40～60g/L时,获得了较高的木糖醇得率,约0.6g/g,但是木糖消耗速率较慢。     

正交旋转回归试验优化木糖醇发酵培养基

利用旋转回归法研究热带假丝酵母 (Candidatropicalis)木糖醇发酵的 2个因素 :葡萄糖和酵母膏用量对木糖醇转化率的影响 ,根据木糖醇转化率依葡萄糖和酵母膏用量的回归方程分析表明 ,培养基中添加葡萄糖能提高木糖醇产物转化率 ,而酵母膏对提高产物转化率的作用不显著。研究同时表明 ,添加葡萄糖后能降低培养基中酵母膏的使用量 ,节约成本。根据回归方程寻优得出 :当木糖质量浓度 13 0 g/L时 ,葡萄糖用量 14 2 6g/L ,酵母膏用量3 40 g/L时 ,由木糖生成木糖醇的产物转化率最高     

Xylitol production on sugarcane biomass hydrolysate by newly identified Candida tropicalis JA2 strain

Xylitol is a building block for a variety of chemical commodities, besides being widely used as a sugar substitute in the food and pharmaceutical industries. The aim of this work was to develop a microbial process for xylitol production using sugarcane bagasse hydrolysate as substrate. In this context, 218 non-Saccharomyces yeast strains were screened by growth on steam-exploded sugarcane bagasse hydrolysate containing a high concentration of acetic acid (8.0 g/L). Seven new Candida tropicalis strains were selected and identified, and their ability to produce xylitol on hydrolysate at low pH (4.6) under aerobic conditions was evaluated. The most efficient strain, designated C. tropicalis JA2, was capable of producing xylitol with a yield of 0.47 g/g of consumed xylose. To improve xylitol production by C. tropicalis JA2, a series of experimental procedures were employed to optimize pH and temperature conditions, as well as nutrient source, and initial xylose and inoculum concentrations. C. tropicalis JA2 was able to produce 109.5 g/L of xylitol with a yield of 0.86 g/g of consumed xylose, and with a productivity of 2.81 g·L·h, on sugarcane bagasse hydrolysate containing 8.0 g/L acetic acid and177 g/L xylose, supplemented with 2.0 g/L yeast nitrogen base and 4.0 g/L urea. Thus, it was possible to identify a new C. tropicalis strain and to optimize the xylitol production process using sugarcane bagasse hydrolysate as a substrate. The xylitol yield on biomass hydrolysate containing a high concentration of acetic acidobtained in here is among the best reported in the literature.     

ACA微囊化酵母细胞利用酒糟水解液发酵木糖醇的初步研究

通过ACA(Chitosan-alginate)微胶囊技术包埋的热带假丝酵母(Candidatropicalis)细胞能有效地发酵酒糟(文中均指丢糟)半纤维素水解液生产木糖醇。在摇瓶条件下,适宜的工艺条件为:初始木糖质量浓度100g/L,发酵液初始pH值6.0,限制性供氧,分段改变摇床转速(0～24h为180r/min,24～48h为120r/min)使菌株在培养早期获得较高水平的通气率,而后降低菌株的呼吸率。每升氮源含酵母膏1.8g,蛋白胨3.0g,微囊化胶珠与水解液体积比为1∶4。此方法有望大幅降低原料预处理的成本,发酵结果良好,显示了良好的应用潜力。     

THE ENHANCEMENT OF AMMONIA PRETREATMENT ON THE FERMENTATION OF RICE STRAW HYDROLYSATE TO XYLITOL

In xylitol fermentation from rice straw hydrolysate by Candida tropicalis As2.1776, ammonia steeping was employed to pretreat the rice straw. Experimental results showed that the content of toxic compounds created in the hydrolysis process, such as acetic acid and phenolic compounds, was greatly reduced and the fermentation of the hydrolysate was enhanced. Xylitol fermentation was investigated in flasks and a 2‐L bioreactor, respectively. The xylitol yield factor and volumetric productivity were 0.746 g/g and 0.686 g/(L·h) in the lab‐flask fermentation, respectively. The corresponding results conducted in bioreactor fermentation were 0.689 g/g and 0.697 g/(L·h), respectively.     

Use of spent brewer's yeast in L‐(+) lactic acid fermentation

The application of by‐products from the brewing industry in lactic acid (LA) production was investigated in order to replace expensive nitrogen sources (such as yeast extract) with cheaper and renewable nitrogenous materials such as brewer's yeast (BY). In this study, brewer's spent grain (BSG) hydrolysate was used for L‐(+)‐LA fermentation by Lactobacillus rhamnosus ATCC 7469. The effect of pH control during the fermentation and the addition of various BY contents (5–50 g/L) in BSG hydrolysate on fermentation parameters was evaluated. BY addition significantly increased free amino nitrogen (FAN) concentration (by 25.2% at 5 g/L to 616% at 50 g/L). A strong positive correlation between FAN concentration in the hydrolysate and concentration of L‐(+)‐LA produced was observed (correlation coefficient of 0.913). A high cell viability of L. rhamnosus ATCC 7469 (1.95–3.32 × 10⁹ CFU/mL at the end of fermentation) was achieved in all fermentations with the addition of brewer's yeast. The addition of BY increased L‐(+)‐lactic acid yield and volumetric productivity up to 8.4% (5 g/L) and 48.3% (50 g/L). The highest L‐(+)‐LA yield (89%) and volumetric productivity (0.89 g/L h^?1) were achieved in fermentation of BSG hydrolysate with 50 g/L of BY. © 2019 The Institute of Brewing & Distilling     

莫格假丝酵母微胶囊化生产木糖醇的研究

研究用高压微胶囊成型装置制备莫格假丝酵母NaCS-PDMDAAC微胶囊生产木糖醇。考察了PDMDAAC浓度对微胶囊性能的影响及推进速度、电压、针头内径对微胶囊直径的影响。摇瓶条件下,微胶囊细胞连续发酵9批,粒径2.0mm的微囊得到最大木糖醇浓度为27.6g/L,发酵时间从游离培养的103h缩短为75h,最大生产能力为0.37g/(L•h),比游离细胞提高9.1%。     

Increase of xylitol productivity by cell-recycle fermentation of Candida tropicalis using submerged membrane bioreactor

Candida tropicalis, an osmophilic strain isolated from honeycomb, produced xylitol at a maximal volumetric productivity of 3.5 g l(-1) h(-1) from an initial xylose concentration of 200 g l(-1). Even at a very high xylose concentration, e.g., 350 g l(-1), this strain produced xylitol at a moderate rate of 2.07 g l(-1) h(-1). In a fed-batch fermentation of xylose and glucose, 260 g l(-1) xylose was added, and the xylitol production was 234 g l(-1) for 48 h, corresponding to a rate of 4.88 g l(-1) h(-1). To increase xylitol productivity, cells were recycled in a submerged membrane bioreactor with suction pressure and air sparging. For each recycle round in cell-recycle fermentation, the average concentration of xylitol produced, fermentation time, volumetric productivity, and product yield were 180 g l(-1), 19.5 h, 8.5 g l(-1) h(-1), and 85%, respectively. When cell-recycle fermentation was started with the cell mass concentrated twofold after batch fermentation and performed for 10 recycle rounds, we achieved a very high productivity of 12 g l(-1) h(-1). The productivity and total amount of xylitol in cell-recycle fermentation were 3.4- and 11.0-fold higher than those in batch fermentation, respectively.     

Fed‐batch l‐(+)‐lactic acid fermentation of brewer's spent grain hydrolysate

Brewer's spent grain (BSG) hydrolysates were used for l ‐(+)‐lactic acid (LA) fermentation by Lactobacillus rhamnosus ATCC 7469. The aim of this study was to evaluate fed‐batch LA fermentation of BSG hydrolysate with the addition of glucose, glucose and yeast extract, and wort during LA fermentation and its effect on fermentation parameters such as LA concentration, its volumetric productivity and yield, and L. rhamnosus cell viability. The highest LA yield, volumetric productivity and concentration of 93.3%, 2.0 g/L/h, and 116.1 g/L, respectively, were achieved with glucose and yeast extract addition during fermentation. In fed‐batch fermentation with glucose and yeast extract addition significantly higher LA concentration, yield and volumetric productivity (by 194.8; 2.2, and 20.7%, respectively) were achieved compared with batch fermentation. The results indicated that fed‐batch fermentation could be used to increase LA fermentation efficiency. Copyright © 2017 The Institute of Brewing & Distilling