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     

新型合金催化剂用于木糖加氢制木糖醇的正交试验研究

采用RTH 311催化剂进行木糖加氢,考察了温度、压力、pH值、剂糖比和反应时间等因素对木糖加氢反应的影响,并采用正交试验方法得出木糖转化率达到最大时的工艺参数。结果表明,当pH值为65、反应温度140 ℃、反应压力10 MPa、反应时间为110 min、剂糖质量比为25%时,木糖转化率高达99.99%。     

新型合金催化剂用于木糖加氢制木糖醇的研究

采用三元Raney Ni合金催化剂进行木糖加氢，考察了温度、压力、pH值、剂糖比和反应时间等因素对加氢反应的影响，并与其它Raney Ni类催化剂进行了对比。结果表明，使用Raney Ni催化剂RTH-311进行木糖加氢，当反应温度为130-140℃、压力为8-10MPa、pH值为6-7、剂糖质量比为2％、反应时间为90～120min时，转化率可达100％，产品色度好。RTH-311催化剂的木糖加氢活性好于其它类催化剂，重复使用时，也可保持较高活性。     

Induction of xylose reductase and xylitol dehydrogenase activities on mixed sugars in Candida guilliermondii

The ability of various sugars to induce xylose reductase (aldose reductase, EC 1.1.1.21) and xylitol dehydrogenase (D -xylulose reductase, EC 1.1.1.9) activities in Candida guilliermondii was studied. D -Xylose was found to be the best inducer of activities of both enzymes, followed closely by L -arabinose. Very low xylose reductase activity was induced by cellobiose, D -mannose, D -glucose, D -galactose, D -fructose or glycerol. With xylitol dehydrogenase, cellobiose and D -fructose caused partial induction of enzyme activity, while negligible activity was induced by D -mannose, D -glucose, D -galactose or glycerol. Several sugars were tested for the ability to repress the induction by D -xylose of xylose reductase and xylitol dehydrogenase activities in C. guilliermondii. Enzyme activities induced on D -xylose served as controls. L -Arabinose, cellobiose and D -galactose did not repress enzyme induction by D -xylose, while D -mannose, D -glucose and D -fructose repressed enzyme induction to varying extents. Results from enzyme induction generally correlated with patterns of mixed sugar utilization, with some anomalies associated with the utilization of D -galactose and D -fructose in the presence of D -xylose. The results show that the utilization of D -xylose by C. guilliermondii is subject to regulation by induction and catabolite repression.     

Improved xylitol production in media containing phenolic aldehydes: application of response surface methodology for optimization and modeling of bioprocess

BACKGROUND: The combined effects of vanillin and syringaldehyde on xylitol production by Candida guilliermondii using response surface methodology (RSM) have been studied. A 2² full‐factorial central composite design was employed for experimental design and analysis of the results. RESULTS: Maximum xylitol productivities (Q_P = 0.74 g L^?1h^?1) and yields (Y_P/S = 0.81 g g^?1) can be attained by adding only vanillin at 2.0 g L^?1 to the fermentation medium. These data were closely correlated with the experimental results obtained (0.69 ± 0.04 g L^?1 h^?1 and 0.77 ± 0.01 g g^?1) indicating a good agreement with the predicted value. C. guilliermondii was able to convert vanillin completely after 24 h of fermentation with 94% yield of vanillyl alcohol. CONCLUSIONS: The bioconversion of xylose into xylitol by C. guilliermondii is strongly dependent on the combination of aldehydes and phenolics in the fermentation medium. Vanillin is a source of phenolic compound able to improve xylitol production by yeast. The conversion of vanillin to alcohol vanilyl reveals the potential of this yeast for medium detoxification. Copyright © 2009 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     

Detoxification of sugarcane bagasse hemicellulosic hydrolysate with ion‐exchange resins for xylitol production by calcium alginate‐entrapped cells

This study describes the performance of four different resins, in sequence, to detoxify sugarcane bagasse hemicellulosic hydrolysate and to improve xylitol production by calcium alginate‐entrapped Candida guilliermondii FTI20037 cells under conditions of low oxygen concentration. The treatment resulted in a removal of 82.1% furfural, 66.5% hydroxymethylfurfural, 61.9% phenolic compounds derived from lignin degradation, 100% chromium, 46.1% zinc, 28.5% iron, 14.7% sodium and 3.5% nickel. On the other hand, the removal of acetic acid was not significant. A xylitol yield factor (Y_P/S) of 0.62 g g^?1 and a volumetric productivity (Q_p) of 0.24 g dm^?3 h^?1 were attained in the fermentation process for xylitol production from detoxified hydrolysate. Copyright © 2004 Society of Chemical Industry     

Identification of optimum synthesis conditions for a novel anion exchange membrane by response surface methodology

The Response Surface Methodology (RSM) was employed for the optimization of the synthesis conditions of an anion exchange membrane. A novel chlorinated‐polypropylene heterogeneous anion exchange membrane was made via phase inversion. A nonionic surfactant was incorporated into the composition as an additive to enhance the membrane properties. The membrane performance was measured in terms of ion exchange capacity (IEC) and permselectivity. An experimental design was used to quantify the effects of variables including the ratio of resin/polymer, the ratio of additive/total solid, and the ratio of solvent/polymer, on IEC and permselectivity. For each function, a quadratic model was developed to correlate the relationship between variables and the response. The results demonstrated the accuracy of the two models. The anion exchange membrane with the best combination of a high IEC and high permselectivity was synthesized with a solvent/polymer ratio of 18.63 (v/w), resin/polymer ratio of 1 (w/w), and additive/total solid ratio of 0.02 (w/w). © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39888.     

Prediction of optimum reaction conditions for the thermo‐tolerant acetylxylan esterase from Neocallimastix patriciarum using the response surface methodology

BACKGROUND: Xylan is the second most abundant renewable polysaccharide in nature and also represents an important industrial substrate. The complete degradation of xylan requires the combination of several types of xylanolytic enzymes, including endo‐β‐1,4‐xylanases, β‐xylosidases, and acetylxylan esterases. As a biocatalyst, xylanolytic enzymes with good thermal stability are of great interest, therefore, a thermo‐tolerant acetylxylan esterase, AxeS20E, was investigated. RESULTS: The cDNA encoding the carbohydrate esterase (CE) domain of AxeS20E from Neocallimastix patriciarum was expressed in Escherichia coli as a recombinant His₆ fusion protein. The recombinant AxeS20E protein was obtained after purification by immobilized metal ion‐affinity chromatography. Response surface modeling (RSM) combined with central composite design (CCD) and regression analysis were then employed for the planned statistical optimization of the acetylxylan esterase activities of AxeS20E. The optimal conditions for the highest activity of AxeS20E were observed at 54.6 °C and pH 7.8. Furthermore, AxeS20E retained more than 85% of its initial activity after 120 min of heating at 80 °C. CONCLUSIONS: The results suggested that RSM combined with CCD and regression analysis were effective in determining optimized temperature and pH conditions for the enzyme activity of AxeS20E. The results also proved AxeS20E was thermo‐tolerant and might be a good candidate for various biotechnological applications. Copyright © 2009 Society of Chemical Industry     

响应面法优化壳聚糖酶产生菌Mitsuaria sp.K1的产酶发酵条件

壳聚糖（chitosan）及其降解产物因具有优良的物理特性和生物活性而被广泛关注。通过平板透明圈初筛、摇瓶复筛方法,从青岛海岸土壤中分离筛选到1株产壳聚糖酶活性较高的细菌Mitsuaria sp.K1,并对其产酶发酵条件进行了单因素试验和响应面优化分析试验。结果表明：在最适培养基组成（1%粉末壳聚糖、0.5%硝酸钾、0.22%KH2PO4、0.1%Na2HPO4、0.15%KCl、0.05%MgSO4?7H2O）和最佳培养条件（培养温度25.2 ℃,培养时间25.4 h,起始pH值6.5,接种量3%,装液量100 mL/500 mL摇瓶,160 r/min）下,Mitsuaria sp.K1的发酵粗酶液最高酶活平均达11.56 U/mL,比优化前的2.17 U/mL提高了4.32倍。与前人研究结果相比,该菌发酵产酶温度降低了5～10 ℃,产酶周期缩短了23～47 h,因此具有工业发酵应用价值。     

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.     

Fermentation optimization for the production of lovastatin by Aspergillus terreus: use of response surface methodology

A Box–Behnken experimental design was used to investigate the effects of five factors—ie oxygen content in the gas phase; concentrations of C, N and P; and fermentation time—on the concentrations of biomass and lovastatin produced in batch cultures of Aspergillus terreus. The values of the various factors in the experiment ranged widely, as follows: 20–80% (v/v) oxygen in the aeration gas; 8–48 g dm^?3 C‐concentration; 0.2–0.6 g dm^?3 N‐concentration; 0.5–2.5 g dm^?3 phosphate‐concentration; and 7–11 days fermentation time. No previous work has used statistical analysis in documenting the interactions between oxygen supply and nutrient concentrations in lovastatin production. The Box–Behnken design identified the oxygen content in the gas phase as the principal factor influencing the production of lovastatin. Both a limitation and excess of oxygen reduced lovastatin titers. A medium containing 48 g dm^?3 C supplied as lactose, 0.46 g dm^?3 N supplied as soybean meal, and 0.79 g dm^?3 phosphate supplied as KH₂PO₄, was shown to support high titers (～230 mg dm^?3) of lovastatin in a 7‐day fermentation in oxygen‐rich conditions (80% v/v oxygen in the aeration gas). Under these conditions, the culture medium had excess carbon but limiting amounts of nitrogen. The optimized fermentation conditions raised the lovastatin titer by four‐fold compared with the worst‐case scenario within the range of factors investigated. Copyright © 2004 Society of Chemical Industry     

A study on the recovery of xylitol by batch adsorption and crystallization from fermented sugarcane bagasse hydrolysate

Xylitol was recovered from fermented sugarcane bagasse hemicellulosic hydrolysate by adsorption and crystallization procedures. Silica gel adsorption was employed to purify the broth containing xylitol. In this step, different mixtures of the solvents ethyl acetate, ethanol and acetone were used as eluent, and different proportions of fermented broth volume incorporated per gram of silica gel (V_b/M_sg, varying from 1.0 to 2.0 cm³ g^?1) were used to pack the column employed as stationary phase bed. The xylitol purification efficiency varied for each mixture of solvent, and for each V_b/M_sg ratio used. The purified broth was submitted to different crystallization procedures (cooling, concentration and supplementation with commercial xylitol) aiming to recover xylitol crystals. The best result (60% crystallization yield and 33% total recovery of xylitol from fermented broth) was obtained when the column was packed with a V_b/M_sg ratio of 2 cm³ g^?1, and the broth was purified with a mixture of ethyl acetate and ethanol, concentrated 6.5‐fold, and supplemented with commercial xylitol to force the precipitation. Copyright © 2006 Society of Chemical Industry     

Comparison of different procedures for the detoxification of eucalyptus hemicellulosic hydrolysate for use in fermentative processes

Eucalyptus (Eucalyptus grandis) shavings were submitted to an acid hydrolysis process with the aim of obtaining a hemicellulosic hydrolysate rich in fermentable sugars. However, the hydrolysate obtained contained, in addition to sugars, several compounds that are toxic to microorganisms, namely furfural, hydroxymethylfurfural, acetic acid and phenolics. In order to produce a hydrolysate suitable for use in fermentative processes, several procedures were evaluated for hydrolysate detoxification, including concentration by vacuum evaporation and adsorption on activated charcoal, diatomaceous earths, ion‐exchange resin or adsorbent resin. Hydrolysate concentration was especially effective for furfural removal, whereas the adsorbent resin was efficient in removing hydroxymethylfurfural, phenolics and acetic acid. Combination of this resin with activated charcoal was better than with diatomaceous earths for removal of acetic acid and phenolics. The best detoxification procedure evaluated was based on hydrolysate concentration followed by adsorption on activated charcoal and adsorbent resin. By this treatment, removal rates of 82.5, 100, 100 and 94% were attained for acetic acid, furfural, hydroxymethylfurfural and phenolics, respectively. Copyright © 2005 Society of Chemical Industry     

响应面法优化一水硫酸氢钠流化催化精馏生产乙酸乙酯工艺条件

以新型的一水硫酸氢钠为催化剂,采用粉末状催化剂混合乙酸一起进料的加入方式,即流化催化精馏工艺,并利用响应面法优化乙酸乙酯催化精馏过程条件.首先,通过单因素灵敏度分析法对乙酸进料量、酸醇进料摩尔比、回流比、催化剂用量、釜底加热功率5个因素进行实验考察,确定了乙酸进料量、酸醇摩尔进料比、回流比3个关键因素的优化值及取值范围.根据单因素实验结果与精馏塔设备要求,塔釜加热功率和催化剂用量分别设定为68 W和2.0%(质量分数)乙酸,采用中心组合设计原则对乙酸进料量、酸醇进料摩尔比和回流比3个关键因素进行实验设计.以乙醇转化率为响应值,基于响应实验结果,利用响应面法对实验结果进行了方程回归,得到3个关键因素与响应值的二次关联模型.通过方差分析和平行实验,证明该模型准确可用.优化后的乙酸乙酯流态化催化精馏工艺条件为乙酸进料量3.2 mol·h^-1,酸醇进料摩尔比为3.1,回流比为3.3,在此优化条件下进行实验,乙醇转化率为88.67%,比基于单因素灵敏度分析法得到的优化工艺条件下乙醇转化率高1.0%.     

响应面分析法优化生姜挥发油提取工艺

在单因素实验的基础上,采用中心组合实验设计及响应面分析法优化了生姜中挥发油的水蒸汽蒸馏提取工艺。结果表明,提取时间(p<0.01),料液比(p<0.05)对挥发油提取得率具有显著影响。最佳工艺条件为:生姜干燥后粉碎过60目筛,加水15.6倍,水蒸汽蒸馏4.78 h。在此条件下,挥发油得率为2.00%,实验结果与模型预测值相符。     

Statistical optimization of process parameters for the production of vanillic acid by solid‐state fermentation of groundnut shell waste using response surface methodology

BACKGROUND: Vanillic acid is a flavoring agent and also serves as precursor for vanillin production. Culture medium and fermentation condition for the single step production of vanillic acid from Phanerochaete chrysosporium using lignocellulosic waste as a substrate under solid state fermentation (SSF) were optimized using response surface methodology. RESULTS: The process parameters were chosen by borrowing methodology, and L‐asparagine, pH and moisture content of the solid medium during SSF were identified as the most significant variables. The optimum value of selected variable and their mutual interactions were determined by response surface methodology. The result demonstrated that a yield of 73.58 mg vanillic acid g^?1 substate was predicted under optimum conditions (L‐asparagine 5.98 mmol L^?1 (2.37 mg g^?1 groundnut shell), pH of solid medium 4.51 and moisture content 74.83%). The predicted response was experimentally validated and resulted in a maximum vanillic acid yield of 73.69 mg g^?1 after 8 days of SSF. CONCLUSION: The optimization of fermentation variables resulted in a maximum 10‐fold increase in vanillic acid yield compared with that observed under sub‐optimal conditions (from 7.2 mg g^?1 to 73.69 mg g^?1). Copyright © 2011 Society of Chemical Industry