Effect of surfactants on separate hydrolysis fermentation and simultaneous saccharification fermentation of pretreated lodgepole pine

The effects of surfactants addition on enzymatic hydrolysis and subsequent fermentation of steam exploded lodgepole pine (SELP) and ethanol pretreated lodgepole pine (EPLP) were investigated in this study. Supplementing Tween 80 during cellulase hydrolysis of SELP resulted in a 32% increase in the cellulose‐to‐glucose yield. However, little improvement was obtained from hydrolyzing EPLP in the presence of the same amount of surfactant. The positive effect of surfactants on SELP hydrolysis led to an increase in final ethanol yield after the fermentation. It was found that the addition of surfactant led to a substantial increase in the amount of free enzymes in the 48 h hydrolysates derived from both substrates. The effect of surfactant addition on final ethanol yield of simultaneous saccharification and fermentation (SSF) was also investigated by using SELP in the presence of additional furfural and hydroxymethylfurfural (HMF). The results showed that the surfactants slightly increased the conversion rates of furfural and HMF during SSF process by Saccharomyces cerevisiae. The presence of furfural and HMF at the experimental concentrations did not affect the final ethanol concentration either. The strategy of applying surfactants in cellulase recycling to reduce enzyme cost is presented. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

丝状真菌Amorphotheca resinae ZN1的糠醛降解代谢分析

木质纤维素在预处理过程产生的降解产物对后续的酶水解和微生物发酵过程产生了强烈的抑制。因此,这些抑制物的脱除即所谓的"脱毒"步骤是正常进行后续酶解和发酵的前提条件。我们对本实验室筛选的丝状真菌Amorphotheca resinae ZN1的糠醛的代谢路径进行了研究。丝状真菌A.resinae ZN1转化糠醛的降解代谢途径可以简述为:糠醛首先快速地转化为毒性较低的糠醇;在有氧条件下,糠醇又再度生成不致对微生物产生危害的低浓度糠醛,糠醛继续氧化为糠酸。推测糠酸可能继续进入TCA循环,进而完成糠醛的完全降解。研究结果为将来加快丝状真菌A.resinae ZN1生物脱毒速率、改善木质纤维素生物转化的限速步骤提供了重要的实验依据。     

Furfural, 5-hydroxymethyl furfural, and acetoin act as external electron acceptors during anaerobic fermentation of xylose in recombinant Saccharomyces cerevisiae

The electron acceptors acetoin, acetaldehyde, furfural, and 5-hydroxymethylfurfural (HMF) were added to anaerobic batch fermentation of xylose by recombinant, xylose utilising Saccharomyces cerevisiae TMB 3001. The intracellular fluxes during xylose fermentation before and after acetoin addition were calculated with metabolic flux analysis. Acetoin halted xylitol excretion and decreased the flux through the oxidative pentose phosphate pathway. The yield of ethanol increased from 0.62 mol ethanol/mol xylose to 1.35 mol ethanol/mol xylose, and the cell more than doubled its specific ATP production after acetoin addition compared to fermentation of xylose only. This did, however, not result in biomass growth. The xylitol excretion was also decreased by furfural and acetaldehyde but was unchanged by HMF. Thus, furfural present in lignocellulosic hydrolysate can be beneficial for ethanolic fermentation of xylose. Enzymatic analyses showed that the reduction of acetoin and furfural required NADH, whereas the reduction of HMF required NADPH. The enzymatic activity responsible for furfural reduction was considerably higher than for HMF reduction and also in situ furfural conversion was higher than HMF conversion.     

糠醛和5-羟甲基糠醛对大肠杆菌产丁二酸的影响

利用可再生生物质特别是木质纤维素水解液来生产平台化合物丁二酸,是目前研究的热点。虽然许多研究者相继报道了木质纤维素水解液对菌株生长和丁二酸生产存在一定抑制作用,但并没有水解液中各种抑制物对菌株影响的相关动力学研究及机理研究。我们选择了两种代表性木质纤维素水解液抑制物,即糠醛和5-羟甲基糠醛,系统研究了它们对大肠杆菌的生长和丁二酸生产的影响。结果表明：糠醛和5-羟甲基糠醛的初始抑制浓度均为0.8 g/L。当糠醛浓度大于6.4 g/L,5-羟甲基糠醛浓度大于12.8 g/L时,菌株生长完全受到抑制。在最高耐受浓度下,糠醛的存在使菌株生物量比对照菌株下降77.8%,丁二酸产量下降36.1%。5-羟甲基糠醛的存在使菌株生物量比对照菌株降低13.6%,丁二酸产量降低18.3%。糠醛和5-羟甲基糠醛具有明显的协同作用。体外酶活测定表明丁二酸生产途径中关键酶磷酸烯醇式丙酮酸羧化酶、苹果酸脱氢酶、富马酸还原酶均受糠醛和5-羟甲基糠醛抑制。研究结果对丁二酸生产用纤维素水解液的预处理和脱毒工艺开发具有指导作用,有利于实现丁二酸发酵生产的工业化。     

分别利用产甘油假丝酵母抗逆转录因子CgSTB5和CgSEF1对酿酒酵母菌株糠醛耐受改造

【背景】纤维素是生物转化解决能源问题的主要原料之一,其水解物中存在严重影响抑制菌株生长的糠醛,需脱毒才可应用于发酵,提高菌株耐受性是解决纤维素水解液实际生产应用的关键。【目的】酿酒酵母(Saccharomyces cerevisiae)是主要的纤维素水解液发酵工业菌株,但糠醛耐受性较低,通过分子改造获得具有高糠醛耐受性的菌株。【方法】利用新获得的产甘油假丝酵母(Candidaglycerinogenes)的相关抗逆转录因子CgSTB5、CgSEF1和CgCAS5,通过分子技术进行S.cerevisiae改造,考察其对酿酒酵母糠醛耐受性的影响,并尝试应用于未脱毒纤维素乙醇发酵。【结果】单个表达CgSTB5和CgSEF1的酿酒酵母,通过菌株点板实验表明菌株的糠醛耐受性提高25%以上,并且摇瓶发酵结果显示糠醛降解性能明显提高,生长延滞期明显缩短,S.cerevisiae W303/p414-CgSTB5的未脱毒纤维素乙醇发酵生产效率提高12.5%左右。【结论】转录因子CgSTB5和CgSEF1均能对提高酿酒酵母糠醛耐受性起到重要作用,并且有助于提高酿酒酵母菌株未脱毒纤维素乙醇发酵性能。     

Intracellular acidification as a mechanism for the inhibition by acid hydrolysis-derived inhibitors of xylose fermentation by yeasts

The main degradation products (furfural, hydroxymethylfurfural, acetate) derived from acid hydrolysis of hemicellulosic materials inhibit growth on and fermentation of xylose by Pachysolen tannophilus and Pichia stipitis, with the latter yeast being the more sensitive. The inhibitory effect was more severe when the inhibitors were present together in the medium. Agarose immobilization partially protected the yeasts from the deleterious effects of these compounds. Intracellular de-energization and acidification may be the mechanism by which these compounds exert their toxic effect on the yeast cells. Received 22 July 1997/ Accepted in revised form 11 November 1997     

Isolation of a plasma protein observed in patients with essential hypertension

Daily progesterone administration (1.33 mg/kg body weight) to immature rabbits brought about an initial increase in the uterine content of uteroglobin which, however, subsided when progesterone treatment was continued for 10 days. During this treatment period progesterone did not lose its own uterine receptors nor did it lose its inhibitory effect on the accumulation of occupied nuclear estrogen receptors. Since immature rabbits were used, the decrease of uteroglobin concentration cannot be explained by inhibitory effects of endogenous estrogens. The results suggest that termination of uteroglobin secretion may be a selective and inherent effect of progesterone itself.     

Changes in the populations of microorganisms associated with the application of soil amendments to control Sclerotium rolfsii Sacc.

Populations of microorganisms from soil treated with guanidine thiocyanate, guanylurea sulfate, thiourea, or furfural were compared with those of untreated soil. The materials effected quantitative and/or qualitative changes in composition of the soil microflora depending on the compound used. Guanidine thiocyanate (Gt) significantly (p0.05) increased total fungal populations relative to populations of other treatments. Populations of Penicillium purpurogenum were markedly higher in Gt-treated soil. Gt also increased total bacterial populations, and was the only compound that increased actinomycete populations. The relative percentage of Trichoderma harzianum was significantly higher in soil treated with thiourea than in the other treatments. Furfural increased the percentage of P. purpurogenum with respect to total fungi, and was as effective as guanylurea sulfate in increasing chitinolytic bacteria and those in the Pseudomonas cepacia-group. Thiourea most effectively promoted proliferation of coryneform bacteria. Chitinolytic fungi increased synergistically when Gt and guanylurea sulfate were applied in combination.