细菌产木聚糖酶发酵条件的研究*

研究了碳源、氮源以及其他因子对木聚糖酶高产菌ＷＬＵＮ０２４（Ｐｓｅｕｄｏｍｏｎａｓ ｓｐ．）产酶的影响,结果表明在麸皮６ｇ／Ｌ、（ＮＨ４）２ＳＯ４ ０．８ｇ／Ｌ、Ｋ２ＨＰＯ４ ０．４ｇ／Ｌ、接种量５％－１０％的条件下,３７℃培养３６ｈ,其木聚糖酶活力可达６００ＩＵ／ｍＬ。同时研究了在较优条件下该菌的摇瓶产酶曲线。     

纤维素分解混合菌的扩繁条件研究

目的:开发纤维素分解混合菌制剂。方法:以菌数、纤维素酶活性及木聚糖酶活性做为评价指标,对具有开发潜力的菌群进行了培养条件研究。结果:该菌群的最适培养温度为35℃;最适接种量为1%;培养基的初始pH值以7.0-7.5为好;在培养基中添加一定量的FeSO4,有利于菌的生长;适当控制氧气浓度,可以确保混合菌群中纤维素酶产生菌的正常繁殖。结论:获得了纤维素分解混合菌的扩繁条件。     

不同培养条件下纤维素分解菌对稻草的分解研究

通过不同培养条件 4株纤维素分解菌对稻草的分解试验 ,发现不同纤维素分解菌对稻草纤维素的分解能力有一定的差异 ,部分菌株混合接种培养的分解率明显高于单独培养的分解率 ,混合培养时分解率受接种顺序影响不明显 ,在 1株分解木质素较强菌株 (侧孢霉 )存在情况下 ,培养前期升高培养温度能提高分解率     

桑粒肩天牛肠道纤维素分解细菌的分离和鉴定*

为研究天牛消化利用纤维素的机理,利用纤维素-刚果红琼脂培养基,从桑粒肩天牛(A.germari)幼虫中肠分离到一种兼性厌氧纤维素分解菌。菌落为白色圆形,边缘比较规则,周围溶纤维素透明圈直径一般可达10mm～20mm,细菌大小为0.5～0.8μm×1～3μm,革兰氏染色阳性,杆状,极生鞭毛,无芽孢。结合生化实验结果,初步鉴定该菌株为纤维单孢菌属(Cellulomonas)。     

整肠生菌的生长条件和罐上发酵试验*

对整肠生菌进行了最佳生长条件的研究 ,获得了该菌生长的最佳C/N为 4～ 4 5∶1 ,以及所需的各种维生素、氨基酸 ,提出了优化的生长培养基组成。经发酵中试表明 ,在最适pH 5 5～ 6 5 ,最适生长温度 35℃～ 37℃条件下 ,提高菌数产量 3倍。     

纤维素分解菌对不同纤维素类物质的分解作用

经过CMC平板、滤纸液化和摇瓶培养试验 ,发现 6株菌中 ,产黄纤维单胞菌 (CellulomonasFlav igena)和康氏木霉 (Trichodermakonigii)分解纤维素类物质的能力比较强 ,对来源不同的纤维素类物质分解能力差异很大 ;真菌与细菌一起接种时 ,分解纤维素类物质的速度明显高于其中任何一个单一菌株 ,说明纤维素类物质的分解需要多种微生物的联合作用     

纤维素分解混合菌扩繁培养基的研究

【摘 要】 以菌数、纤维素酶活性及木聚糖酶活性做为评价指标,进行的单因素和正交试验研究表明：纤维素分解菌群的最佳培养基配方为酵母膏0.5%,（NH4）2SO4 0.5%,淀粉1.5%,FeSO4?7H2O 0.02%,NaCl 6 g,KH2PO4 0.5 g,CaCl2 0.1 g,K2HPO4 0.5 g,CMC-Na 5 g,pH 7.0～7.5。     

复合菌株发酵猪血粉的条件研究*

利用筛选出的2株高产蛋白酶的米曲霉(Aspergillus oryzae)菌株AS100、AS102作猪血粉发酵的主发酵菌株,配以1株酵母菌(Saocharomyces cereisiae)菌株Y113和1株细菌(Bacilus sp．)菌株Asp007为辅助菌株,研究了这四株菌的产酶性能,同时确定了它们在发酵血粉过程中的一系列参数。通过猪血粉发酵,获得了一种高蛋白发酵饲料,其香味浓郁,蛋白质含量高达69％,且富含游离氨基酸,VitD3、烟酸等维生素及Fe等无机元素,可作为禽畜高蛋白源或饲料添加剂。     

曲霉液体发酵产原果胶酶的条件优化研究*

研究了碳源、氮源、金属离子及表面活性剂等对菌株(Aspergillus sp.)XZ-131产原果胶酶的影响.果胶类物质是该菌株产原果胶酶所必需的诱导物.以(NH4)2SO4和(NH4)2HPO4作为氮源时,产酶较高,达到300 U/mL.钙离子及Tween-20均能促进该酶的产生.通过正交试验优化得出该菌株产酶的最佳培养基配方为桔皮粉 1g,(NH4)2SO4 2g,CaCl2 0.015g,Tween-20 0.2mL,KH2PO4 3.8g,K2HPO4*3H2O 0.2g,水 100mL,pH 6.5.     

利用光合菌发酵对玉米秸杆进行转化的研究*

系统研究了光合菌在氨法处理和非氨处理两种环境下,在厌氧、好氧、以及自然条件下对玉米秸秆的转化。通过发酵液中还原糖与蛋白质浓度的测定结果的比较、判断、优选出一种最适合条件下光合菌对玉米转化的途径。研究表明,在以氨化法处理的玉米秸秆为底物与光合菌的发酵实验中,发酵液中的还原糖和蛋白质的尝试都要比非氨法条件下玉米秸秆为底物与光合菌发酵实验中的发酵液中的不原糖和蛋白质的浓度高。实验结果证明了转化产生的还原糖、蛋白质都是光合菌能利用的营养成分,由此达到利用光合菌转化玉米秸秆的研究目的。     

烟叶主要生物酶活性变化规律及其在烟叶发酵中的应用

烟叶中的生物酶在烟叶的生长及发酵过程中起着重要作用.他们能够促进烟叶内部有机物质的分解与转化,调整烟叶中各种化学成分的比例,提高烟叶中香气成分的形成和积累,从而提高烟叶的综合品质.综述了烟叶生长、加工过程中淀粉酶、蛋白酶、多酚氧化酶的活性变化规律,以及外加生物酶在烟叶发酵中的应用现状.     

不同精粗比底物下瘤胃真菌和纤维降解细菌共培养发酵特性及菌群变化

采用体外厌氧共培养技术,研究了瘤胃真菌和纤维降解细菌在不同精粗比(A组为全粗料,B组3∶7,C组5∶5,D组7∶3,E组为全精料)底物下菌群变化及其共培养发酵特性。结果表明:与0h相比,发酵至24h时B组和C组的厌氧真菌数量有较大幅度的上升,A组和D组则有所下降,E组未检测到真菌生长;纤维降解细菌随精粗比的增加呈上升趋势。发酵至48h时,各组均未检测到真菌生长;从A组到C组细菌数量呈上升趋势,此后急剧下降。DGGE结果表明,A、B和C组(精粗比低于5∶5)的DGGE图谱相似,有11条共有条带,但是当精粗比上升到7:3时,条带数目显著下降。随精料比例的增加,整个发酵期共培养系统中pH值显著下降(P<0.05)。整个发酵期间,共培养系统发酵产生的VFA主要为乙酸,丙酸和丁酸的量较少,乙酸与丙酸比值从A组到C组呈下降趋势,此后呈上升趋势。随精料比例的上升,发酵48h时总挥发性脂肪酸浓度从A组到C组呈上升趋势,此后呈下降趋势。发酵48h的羧甲基纤维素酶活和木聚糖酶活均以A组最高,而α-淀粉酶活从A组到D组逐渐增大,而E组最低,仅为B、C、D组的1/4~1/3。     

A serpin in the cellulosome of the anaerobic fungus Piromyces sp. strain E2

A gene encoding a novel component of the cellulolytic complex (cellulosome) of the anaerobic fungus Piromyces sp. strain E2 was identified. The encoded 538 amino acid protein, named celpin, consists of a signal peptide, a positively charged domain of unknown function followed by two fungal dockerins, typical for components of the extracellular fungal cellulosome. The C-terminal end consists of a 380 amino acid serine proteinase inhibitor (or serpin) domain homologue, sharing 30 % identity and 50 % similarity to vertebrate and bacterial serpins. Detailed protein sequence analysis of the serpin domain revealed that it contained all features of a functional serpin. It possesses the conserved amino acids present in more than 70 % of known serpins, and it contained the consensus of inhibiting serpins. Because of the confined space of the fungal cellulosome inside plant tissue and the auto-proteolysis of plant material in the rumen, the fungal serpin is presumably involved in protection of the cellulosome against plant proteinases. The celpin protein of Piromyces sp. strain E2 is the first non-structural, non-hydrolytic fungal cellulosome component. Furthermore, the celpin protein of Piromyces sp. strain E2 is the first representative of a serine proteinase inhibitor of the fungal kingdom.     

A novel multifunctional cellulolytic enzyme screened from metagenomic resources representing ruminal bacteria

Metagenomic resources representing ruminal bacteria were screened for novel exocellulases using a robotic, high-throughput screening system, the novel CelEx-BR12 gene was identified and the predicted CelEx-BR12 protein was characterized. The CelEx-BR12 gene had an open reading frame (ORF) of 1140 base pairs that encoded a 380-amino-acid-protein with a predicted molecular mass of 41.8 kDa. The amino acid sequence was 83% identical to that of a family 5 glycosyl hydrolase from Prevotella ruminicola 23. Codon-optimized CelEx-BR12 was overexpressed in Escherichia coli and purified using Ni–NTA affinity chromatography. The Michaelis–Menten constant (K_m value) and maximal reaction velocity (V_max values) for exocellulase activity were 12.92 μM and 1.55 × 10⁻⁴ μmol min⁻¹, respectively, and the enzyme was optimally active at pH 5.0 and 37 °C. Multifunctional activities were observed against fluorogenic and natural glycosides, such as 4-methylumbelliferyl-β-d-cellobioside (0.3 U mg⁻¹), CMC (105.9 U mg⁻¹), birch wood xylan (132.3 U mg⁻¹), oat spelt xylan (67.9 U mg⁻¹), and 2-hydroxyethyl-cellulose (26.3 U mg⁻¹). Based on these findings, we believe that CelEx-BR12 is an efficient multifunctional enzyme as endocellulase/exocellulase/xylanase activities that may prove useful for biotechnological applications.     

Optimization of fermentation conditions for thermostable cellulase production byThielavia terrestris

Summary Of the eighteen different carbon sources, solka floc was optimal for the induction of cellulases by the thermophilic fungusThielavia terrestris. The temperature optimum for growth was between 44–52°C. The effect of initial and controlled pH on fungal growth and cellulase production was investigated and the results obtained showed that the maximum volumetric productivity (6.07 I.U./1 per h) of filter paper activity was achieved when the pH was controlled at 4.5–5.0.     

棉秸秆降解高温菌株的筛选及产酶分析

从新疆地区分离具有降解棉秸秆纤维素功能的菌株,得到4株耐高温真菌(50°C)。纤维素酶学性质分析表明,该4株菌的纤维素酶具有良好的耐酸性(最适pH为4.5)和耐高温性(最高达60°C)。以羧甲基纤维素钠(CMC-Na)、微结晶纤维素、棉花、滤纸、淀粉、果胶为底物测定酶活力,滤纸酶活力(FPA)最高达2.63 U/mL、淀粉酶活力最高达6.17 U/mL、果胶酶活力最高达5.86 U/mL。4株真菌酶学特性分析表明,该系列菌株在秸秆生物质利用方面有很大的应用潜力。     

Induction and preliminary characterization of intracellular β-glucosidases from a cellulolytic Streptomyces strain

Abstract The cellulolytic actinomycete Streptomyces sp. QM-B814 posasses an intracellular β-glucosidase system which is induced by cellobiose and carboxymethylcellulose. Maximal β-glucosidase activity was attained 8–10 h after inducer addition to exponential phase growing cultures. The induction is depressed in the presence of glucose. The system is composed of two electrophoretically different β-glucosidase forms showing relative molecular masses of about 60 and 35 kDa, and p I values in the range 4.2–4.5. Both β-glucosidases are synthesized de novo. The enzymes share substrate preference and are both inhibited by δ-gluconolactone and p -chloromercuribenzoate. The induction pattern and glucose inhibition are similar for both enzymes.     

食用菌深层发酵的研究进展

发酵工程是现代生物工程技术之一,目前,它在食用菌生产中已得到深入研究和广泛应用。通过回顾食用菌液体深层发酵的研究历史,概述了有关食用菌的液体发酵研究状况,并展望了该技术在我国食用菌生产中的应用前景。     

Involvement of lignocellulolytic enzymes in the decomposition of leaf litter in a subtropical forest

The involvement of ligninolytic and cellulolytic enzymes, such as laccase, lignin peroxidase, manganese peroxidase, carboxymethylcellulase (CMCase), and filter paper activity (FPA), in the decomposition process of leaf litter driven by 6 soil-inhabiting fungi imperfecti was studied under solid-state fermentations. All the tested fungi exhibited varied production profiles of lignocellulolytic enzymes and each caused different losses in total organic matter (TOM) during decomposition. Based on the results, the 6 fungi could be divided into 2 functional groups: Group 1 includes Alternaria sp., Penicillium sp., Acremonium sp., and Trichoderma sp., and Group 2 includes Pestalotiopsis sp. and Aspergillus fumigatus. Group 1, with higher CMCase and FPA activities, showed a higher decomposition rate than the fungi of Group 2 over the first 16 d, and thereafter the cellulolytic activities and decomposition rate slowed down. Group 2 showed the maximum and significantly higher CMCase and FPA activities than those of the Group 1 fungi during the later days. This, combined with the much higher laccase activity, produced a synergistic reaction that led to a much faster average mass loss rate. These results suggest that the fungi of Group 1 are efficient decomposers of cellulose and that the fungi of Group 2 are efficient decomposers of lignocellulose. During cultivation, Pestalotiopsis sp. produced an appreciable amount of laccase activity (0.56+/-0.09 U/ml) without the addition of inducers and caused a loss in TOM of 38.2%+/-3.0%, suggesting that it has high potential to be a new efficient laccase-producing fungus.