Strategies for reducing supplemental medium cost in bioethanol production from waste house wood hydrolysate by ethanologenic Escherichia coli: inoculum size increase and coculture with Saccharomyces cerevisiae

In this paper, we report a simultaneous realization of both efficient ethanol production and saving medium nutrient (corn steep liquor [CSL]) during bioethanol fermentation of overliming-treated hydrolysate of waste house wood (WHW) using ethanologenic Escherichia coli KO11. In cultivation using WHW hydrolysate supplemented with 4% (v/v) CSL and 0.2 g-dry cell weight (DCW)/l E. coli KO11 cells, the overall ethanol yield reached 84% of the theoretical value at 61 h. When we conducted the cultivation with 1% CSL to reduce the supplemental medium cost, the overall ethanol yield remained in the range of 66-72% even at 90 h. We proposed two alternative methods for increasing the overall yield even with 1% CSL. The first method involved increasing the inoculum size of E. coli KO11 up to 0.8 g-DCW/l, where 83% of the overall yield was attained at 60 h of cultivation. The second method involved the coculture of 0.2 g-DCW/l E. coli KO11 together with 0.02 g-DCW/l of Saccharomyces cerevisiae TJ1, and the overall yield reached 81% at 47 h of cultivation.     

Biological detoxification of waste house wood hydrolysate using Ureibacillus thermosphaericus for bioethanol production

Hydrolysates of lignocelluloses hydrolyzed by diluted sulfuric acid contain toxic compounds that inhibit ethanol production by Saccharomyces cerevisiae and the ethanologenic recombinant Escherichia coli KO11. We investigated the biological detoxification of a hydrolysate of waste house wood (WHW) by a thermophilic bacterium, Ureibacillus thermosphaericus. When the hydrolysate was treated with this bacterium at 50 degrees C for 24 h, the ethanol production rate by S. cerevisiae increased markedly and was comparable to that for the hydrolysate treated with an excess amount of calcium hydroxide (overliming). Chromatographic analysis of synthetic hydrolysates containing furfural or 5-hydroxymethyl furfural that are considered to be major toxic compounds in hydrolysates revealed that U. thermosphaericus degrades these compounds. In the WHW hydrolysates, however, the concentrations of these compounds were not decreased markedly by the bacterium. These results suggest that the bacterium degrades minor but more toxic compounds or phenolic compounds in the WHW hydrolysates. The combination of bacterial and overliming treatments of hydrolysates minimized significantly the decrease in ethanol production rate by E. coli KO11 as fermentation proceeded. Because the bacterium grows rapidly and does not consume sugars, our biological detoxification should be useful for bioethanol production from acid hydrolysates of lignocelluloses.     

重组大肠杆菌KO11利用单糖发酵乙醇条件优化

对重组大肠杆菌KO11利用单糖转化乙醇的能力进行了研究与优化,结果表明,KO11对己糖中的半乳糖、甘露糖、葡萄糖和戊糖中的岩藻糖、木糖均有较高的乙醇转化率。通过对KO11利用葡萄糖与木糖发酵的考察发现,以4%(v/v)接种量接种于含葡萄糖20 g/L的LB培养基中,KO11在pH 5.5、37℃、150 r/min振荡培养4 h后再静置发酵20 h能够达到乙醇转化率94.81%;将其接种于含同样浓度木糖的LB培养基在相同的pH、发酵温度与振荡速度下振荡12 h后再静置发酵15 h达到乙醇转化率94.15%。另外,KO11具有较高糖耐受性,发酵葡萄糖与木糖混合糖时优先利用葡萄糖,并且在43℃时仍表现出较高的乙醇转化率,为利用该菌种进行纤维质水解液发酵提供了参考依据。     

Ethanol production from non-pretreated napiergrass through a simultaneous saccharification and fermentation process followed by a pentose fermentation with Escherichia coli KO11

Efficient ethanol production from lignocellulosic napiergrass (Pennisetum purpureum Schumach) was examined by the combination of the simultaneous saccharification and fermentation (SSF) with commercial cellulase and Saccharomyces cerevisiae NBRC 2044 and subsequent pentose fermentation (PF) by Escherichia coli KO11. Under the optimized conditions, the combination of the SSF and PF processes resulted in the production of 144 mg g(-1) of ethanol from the non-pretreated napiergrass powder. The ethanol yield was 44.2% of the theoretical yield based on the hexose (37.5 g) and pentose (26.5 g) derived from 100 g of dry powdered napiergrass.     

某屠宰场大肠杆菌O26的分离与鉴定

为了了解屠宰场大肠杆菌O26污染情况,参考美国农业部(united states department of agriculture,USDA)的检测方法,对样品进行选择性增菌,经免疫磁珠富集、选择性显色培养基mRainbow Agar分离纯化后,挑选可疑菌株采用PCR方法鉴定O抗原,进一步采用血清凝集试验进行验证,并对确认的阳性菌株采用多重PCR方法进行毒力基因(stx1、stx2、eae、hly)检测。结果表明,采集的120份样品中共分离到1株大肠杆菌O26,但上述4种毒力基因均为阴性。该屠宰场存在大肠杆菌O26的污染,但分离出的大肠杆菌O26没有携带毒力基因。     

Disruption of ptsG gene and manXYZ operon of ethanol-producing Escherichia coli KO11: Effects on glucose and xylose utilization and ethanol production

Ethanol-producing Escherichia coli strain KO11 consumed 99% of the glucose and only 13% of the xylose in a mixture of glucose (60g/L) and xylose (40g/L) during the 72-h fermentation at 30°C. The deletion mutants ΔptsG, ΔmanXYZ, and ΔptsG/manXYZ utilized 42%, 78%, and 35% of the glucose and 50%, 32%, and 32% of the xylose, respectively.     

利用重组大肠杆菌发酵生产L-茶氨酸

L-茶氨酸是茶叶中一种独特的非蛋白质氨基酸。γ-谷氨基甲酰胺合成酶可以催化L-谷氨酸、乙胺和ATP合成L-茶氨酸,但反应成本较高。为了提高L-茶氨酸的生产效率,该研究开发了直接发酵生产L-茶氨酸的新方法。首先,在大肠杆菌基因组上双拷贝Methylovorus mays来源的γ-谷氨酰甲胺合成酶基因gmas,获得了一株遗传稳定的用于L-茶氨酸生产的重组菌株。其次,在5 L罐中采用流加乙胺的方式发酵20 h,L-茶氨酸产量达到30. 45 g/L,糖酸转化率达到20.17%。最后,根据L-茶氨酸的物化性质拟定出合适的分离提取路线,从发酵液中获得了98.51%纯度的L-茶氨酸成品,总提取收率可达到70.34%。该方法操作简单,原料成本较低,具有较好的工业应用前景。     

家具生产中木质废料的来源、控制与处理

分析了家具制造企业原料输入和废料产生情况,针对家具生产过程木质废料的产生情况和废料形式进行了剖析,结合生产实际提出了家具生产木质废料的控制与处理措施。     

蒸汽爆破尾叶桉木材纤维素酶水解液发酵单细胞蛋白

用高效液相色谱对蒸汽爆破尾叶桉木材的纤维素酶水解液进行了还原糖和乙酸含量分析,通过酿酒酵母和假丝酵母混合接种,对水解液进行了摇瓶发酵单细胞蛋白的工艺研究,确定了发酵的最佳工艺条件。结果表明,水解液中主要含有葡萄糖和木糖,用双菌种混合发酵单细胞蛋白的最佳发酵条件为:酿酒酵母(Sacchromyces cerevisive):假丝酵母(Candida utilis2.587)为1:3,发酵温度31℃,初始pH4.5,初始还原糖浓度17～18g/L,发酵时间27h。     

废弃纺织品纤维与木浆配抄纸的性能

为探讨废弃纺织品作为原料在现代造纸工业中的应用,按照原料和颜色深浅把废弃纺织品分为4类:合成纤维(浅色)、纤维素纤维(浅色)、纤维素纤维(深色)、纯棉纤维(浅色),将原料经粉碎工序加工成散纤维,分别与木浆配抄成手抄纸,分析了配抄纸的各项物理性能。结果表明,手抄纸具有优异的透气性能,透气度为91.70~100.00μm/(Pa·s),是木浆(对比样)手抄纸的9.6~10.44倍;力学性能低于木浆手抄纸,是木浆手抄纸的27.8%~55.6%;废弃纺织纤维与木浆配抄纸的白度与纺织品纤维原来的颜色相关。     

Quality assessment of pellets and briquettes made from glued wood waste

European Journal of Wood and Wood Products - In accordance with sustainable economic and social development, Europe supports the use of energy from renewable sources to decrease the use of fossil...     

木浆中段废水的脱色处理

为解决木浆中段废水色度难于被生化法所降低的问题，分别采用混凝法，Fenton试剂法，电解法和活性炭吸附法进行了试验研究。     

耐热木聚糖酶基因xyn11EM在大肠杆菌中的表达

将人工合成经密码子优化的11家族耐热木聚糖酶基因xyn11~(EM)克隆至表达质粒p ET-28a(+)中,获得重组质粒p ET-28a-xyn11~(EM)。将其转化Escherichia coli BL21(DE3),构建表达耐热木聚糖酶的重组工程菌E.coli BL21/xyn11~(EM)。用IPTG诱导表达重组木聚糖酶Xyn11~(EM)(re Xyn11~(EM)),酶活性可达47.5 U/m L。SDS-PAGE分析显示,re Xyn11~(EM)的表观相对分子质量为24 800。re Xyn11~(EM)的最适反应温度为70℃,在70℃以下稳定。最适反应p H为6.5~7.0,在p H5.0~8.0范围内稳定。大多数金属离子和EDTA对该重组酶的活性影响不大。re Xyn11~(EM)的Km和Vmax值分别为7.2 mg/m L和54.7 U/mg。结果表明xyn11~(EM)成功在E.coli中实现了异源表达,其良好的热稳定性具有较好的工业应用潜力。     

Inactivation of Escherichia coli on almonds using nonthermal plasma

ABSTRACT:  This study was carried out to investigate the applicability of nonthermal plasma (NTP) technology for the pasteurization of almonds. Almonds were spiked with various levels of Escherichia coli by dipping the almonds in E. coli culture broth followed by drying. The spiked almonds were treated with NTP under different treatment conditions. The pattern of the microorganisms reduction by NTP was analyzed. NTP was found to be effective on reduction of E. coli on almond evidenced by almost 5-log reduction after 30-sec treatment at 30 kV and 2000 Hz. The NTP bactericidal effect on E. coli inoculated on almond increased with the applied voltage and the frequency. The NTP reduction followed the 1st-order reaction kinetics, and the reduction rate constants varied with almond types and grades. The E. coli cells at logarithmic phase were more sensitive to the NTP than those at stationary and declining phases.     

Improvement of productivity of active horseradish peroxidase in Escherichia coli by coexpression of Dsb proteins

Coexpression of two classes of folding accessory proteins, molecular chaperones and foldases, can be expected to improve the productivity of soluble and active recombinant proteins. In this study, horseradish peroxidase (HRP), which has four disulfide bonds, was selected as a model enzyme and overexpressed in Escherichia coli. The effects of coexpression of a series of folding accessory proteins (DnaK, DnaJ, GrpE, GroEL/ES, trigger factor (TF), DsbA, DsbB, DsbC, DsbD, and thioredoxin (Trx)) on the productivity of active HRP in E. coli were examined. Active HRP was produced by very mild induction with 1 microM isopropyl-beta-D-thiogalactopyranoside (IPTG) at 37 degrees C, whereas the amount of active HRP produced by the induction with 1 mM IPTG was negligibly small. Active HRP production was increased significantly by coexpression of DsbA-DsbB (DsbAB) or DsbC-DsbD (DsbCD), while coexpression of molecular chaperones did not improve active HRP production. The growth of E. coli cells was inhibited significantly by the induction with 1 mM IPTG in a HRP single expression system. In contrast, when HRP was coexpressed with DsbCD, the growth inhibition of E. coli was not observed. Therefore, coexpression of Dsb proteins improves both the cell growth and the productivity of HRP.     

Release of CFC-11 from disposal of polyurethane foam waste

The halocarbon CFC-11 has extensively been used as a blowing agent for polyurethane (PUR) insulation foams in home appliances and for residential and industrial construction. Release of CFCs is an important factor in the depletion of the ozone layer. For CFC-11 the future atmospheric concentrations will mainly depend on the continued release from PUR foams. Little is known about rates and time frames of the CFC release from foams especially after treatment and disposal of foam containing waste products. The CFC release is mainly controlled by slow diffusion out through the PUR. From the literature and by reevaluation of an old reported experiment, diffusion coefficients in the range of 0.05-1.7 x 10(-14) m2 s-1 were found reflecting differences in foam properties and experimental designs. Laboratory experiments studying the distribution of CFC in the foam and the short-term releases after shredding showed that about 40% of the CFC is solubilized in the PUR phase, and that up to 10% of the total content will be released within a few weeks if the foam is shredded down to 2-cm sized pieces. For smaller pieces the quick release will be larger. Fifty percent of residual CFC content will be released within 9-300 years from 2-cm pieces based on the range in diffusion coefficients reported. For larger pieces the initial release is insignificant, and the release time frames are much longer than for the shredded foam.     

Analysis of organic solvent tolerance in Escherichia coli using gene expression profiles from DNA microarrays

To investigate the biological mechanism of organic solvent tolerance (OST), DNA microarrays were used to collect and compare the gene expression profiles of normal and organic solvent-tolerant Escherichia coli strains. First, we compared the tolerant-strain OST3410 to its sensitive parent strain JA300 in the absence of organic solvents. Numerous genes showed higher expression levels in OST3410, and Northern analysis was used to confirm the higher expression level of some genes. Next, the gene expression profiles of JA300 and OST3410 exposed to hexane as an organic solvent were investigated and compared with JA300 before exposure to organic solvent. In OST3410 and JA300, 115 and 47 hexane-induced genes were found, respectively. As candidates for genes related to OST, we focused on six genes: cysD, marA, mg1B, tnaA, tnaB and yihM, which were upregulated by hexane in both strains. When these genes were over-expressed on plasmids, only the marA plasmid increased OST activity. It should be noted that we succeeded in finding a gene related to OST activity using only DNA microarray data, without any biochemical or biological knowledge.