外切-β-葡聚糖酶是纤维素酶的重要组分之一,提高该组分的活力是增强纤维素酶协同降解性能、降低纤维素水解成本的关键。分别采用微晶纤维素琼脂平板法和滤纸崩解法,对已有的基因重组转化子进行筛选试验,获得了6个优良转化子,其滤纸崩解速率和微晶纤维素琼脂平板上的生长速率都较大。进一步在摇瓶条件下进行复筛试验,获得了外切-β-葡聚糖酶(C1)高产转化子Trichoderma reesei ZU-101,液体培养48 h,其C1酶活力可达18.24 U·ml-1,是出发菌株的2.16倍;分析结果表明:重组转化子的纤维素酶体系中内切-β-葡聚糖酶和纤维二糖酶的活力与出发菌株相比变化不大,但由于外切-β-葡聚糖酶活力得到了大幅度提高,纤维素酶的总活力(滤纸酶活力FPA)也提高了61.9%。采用纤维素酶对碱预处理玉米秸秆进行酶解试验,当酶用量为20 FPIU·(g底物)-1,水解48 h,重组转化子T.reesei ZU-101纤维素酶的酶解得率高达94.4%。本文的研究结果在可再生纤维素资源的生物转化与利用方面具有广阔的应用前景。

Increasing the activities of exo-β-glucanases, one of the important components of cellulase, is a key to strengthening the synergistic degradation of cellulase and reducing the cost of cellulolytic hydrolysis. The methods of MCC-agar plate screening and liquid culture medium of filter paper were used in this work respectively to screen Trichodermareesei transformants, obtaining 6 superior transformants with higher filter paper collapsing rate and growing rate on MCC-agar plates. Further screening experiment was conducted under shaking flask condition, obtaining the highly exo-β-glucanase (C1)-producing transformant T. reesei ZU-101 whose C1 activity was18.24 U·ml-1 after 48 h liquid culture, 2.16-fold higher than the original strain. Analysis results demonstrated that filter paper activity (FPA) of the cellulase system of the transformant, representing total activity, increased by 61.9%as exo-β-glucanase activity ascended significantly, although endo-β-glucanase and cell

在酸性纤维素酶液中分别加入1%（对溶液质量）的硫脲、硫氰酸钠、亚硫酸钠、亚硫酸氢钠、焦亚硫酸钠、连二亚硫酸钠、硫氢化钠、硫化钠等8种不同的硫化物,在温度40、50、60℃,pH=4.5、5.0、5.5的条件下,根据中心复合试验设计方案测定滤纸酶活力,并用Minitab软件分析各种硫化物对纤维素酶活力的影响.在相同的条件下对纯棉针织物进行酶处理,测定处理前后织物的顶破强力、毛羽去除率、处理残液葡萄糖含量来评价对织物的实际作用效果.结果表明,硫化物的加入可以小幅提高纤维素酶活力、扩大酶的温度和pH作用区间,但除了亚硫酸钠和连二亚硫酸钠外,其他6种硫化物的加入反而降低了织物的除毛效果.

1%(on the mass of acid cellulase solutions) of thiourea, sodium thiocyanate, sodium sulfite, so-dium bisulfite, sodium metabisulfite, sodium hyposulfite, sodium hydrosulfide and sodium sulfide were added to acid cellulase solutions, respectively. The filter paper enzymatic activities were measured according to cen-tral composite experiment design under 40, 50, 60 ℃ and pH 4.5, 5.0, 5.5. The effects of various sulfides on the enzymatic activities were analyzed by Minitab software. The cotton knitted fabrics were treated with cellu-lase under the same conditions, and the bursting strength, hairiness removal percentage and glucose content of residue liquid were measured to evaluate the practical performance on cotton fabrics. The results showed that the addition of sulfides could slightly enhance the cellulase activity, and widen cellulase working tempera-ture and pH range. However, except sodium sulfite and sodium hyposulfite, the addition of other six sulfides decreased the h

为了获得高产纤维素酶菌株,有效地开发和利用纤维素资源.本研究通过对野外采集的大型真菌进行分离纯化,获得了16个菌株.利用CMC固体培养、刚果红染色,测量水解圈与菌落直径的比值(H/C值),对获得的菌株进行初筛;通过液体发酵培养,测定其上清液中的滤纸酶活力(FPA),对菌株进行复筛,最终获得了纤维素酶活性较高的菌株01.以稻草和羧甲基纤维素为碳源,研究了培养温度、pH值、培养时间对真菌菌株01产纤维素酶的影响.结果表明,该菌株产纤维素酶的最适培养温度为27℃,pH值为5.0,培养时间为6 d,菌株01的滤纸酶活性达到580.0 IU/mL.因此,真菌01可作为纤维素酶研究和饲料加工等生产的备选菌株.

Sixteen strains of large fungi collected in the field from Huaihua were isolated and purified . Congo red staining , the hydrolysis circle and colony diameter ratio (H/C value ) were analysed on CMC solid medium to screen cellulase producing fungal strains . By determining the filter paper enzyme activity (FPA ) in its supernatant of liquid fermentation culture , higher cellulase activity strain 01 were obtained ultimately . Factors including culture temperature , pH and incubating time affecting the production of cellulase had been studied by using the rice and CMC as carbon source . The results showed that the best culture temperature was 27℃ , the optimal pH of culture medium and incubating time was 5.0 and 6d , respectively . Under the optimal condition , the FPA reached 580.0IU/mL . In this article , it showed that fungal 01 could been a candidate for future studying cellulase and feed production .

纤维素油脂的统合生物加工过程是将纤维素酶生产、纤维素水解和微生物油脂发酵过程组合,通过一种微生物完成.运用统合生物加工过程生产微生物油脂可以降低生物转化过程的成本.该文对20株纤维素降解菌进行筛选评价,结果发现青霉菌株 P‐2同时具备有效的纤维素降解和油脂积累能力.脂肪酸组分分析表明,菌株 P‐2胞内油脂的脂肪酸组分主要为棕榈酸（ C16：0,21.05％）、油酸（ C18：1,22.43％）和亚油酸（ C18：2,27.78％）.菌株P‐2在以纤维素粉为底物的液体发酵和以秸秆、麸皮混合物为底物的固态发酵条件下可达到的最高油脂产量分别为0.65 g/L 和40.13 mg/g（按干物质计）.说明青霉 P‐2是一株潜在的低成本纤维素油脂生产菌.进一步分析在发酵试验中的油脂产量和纤维素酶活力发现,菌株 P‐2的纤维素酶分泌能力在其油脂生产过程中具有重要作用.外源纤维素酶添加试验证实,在培养基中外源纤维素酶添加量的提高可以促进 P‐2油脂的生产.添加24 IU /g （按干物质计）纤维素酶可使 P‐2发酵后的最高油脂产量达到0.83 g/L .对固态发酵所得到的滤纸酶活力和油脂产量数据进行相关分析,结果证实滤纸酶活力与油脂产量之间存在极显著的正相关关系（ R2＝0.711,P＜0.01）.当固态发酵系

Summary Consolidated bioprocessing (CBP) is a convenient and cost‐efficient strategy to produce single cell oils ( SCOs ) from cellulose‐based substrates . In this study , Penicillium P‐2 having both effective cellulose degradation and lipid accumulation was isolated from 20 cellulolytic fungi . The SCOs of the strain P‐2 were mainly composed of palmitic acid ( C16 :0 , 21 .05% ) , oleic acid ( C18 :1 , 22 .43% ) and linoleic acid ( C18 :2 , 27 .78% ) . Fermentation experiments of the strain P‐2 showed that maximum lipid yields of 0 .65 g/L and 40 .13 mg/g ( per gram dry mass of initial solid substrate) could be obtained by submerged fermentation ( SmF) from cellulose and solid‐state fermentation (SSF) from wheat straw and bran mixture , respectively . The results from the two CBP indicated that the strain P‐2 had a potential to be a promising low‐cost oil producer using cellulose‐based substrates . The further analysis for lipid accumulation and cellulase secretion

木聚糖酶在饲料、食品、纺织、造纸等行业应用广泛,是重要的工业用酶之一。利用丝状真菌—好食脉孢霉(N.sitopHila)为研究对象,以蛋白胨、豆渣、麸皮水和豆粕为底物进行液态发酵生产木聚糖酶,发酵后用滤纸真空抽滤后得粗酶液。用DNS法对脉孢霉半纤维素酶活力进行测定,对脉孢霉产木聚糖酶最优发酵培养基、接种量和培养温度进行研究,研究表明好食脉孢霉产半纤维素酶最适发酵培养基为3%麸皮水、0.5%蛋白胨、6%豆渣、pH自然,最适接种量为1%,最适发酵温度为28℃。

As one of the most important enzymes, xylanase has been widely applied in the fields of feed, food, textile and paper industries. The filiform fungus-neurospora sitophila was used as object of study, and xylanase was produced with using peptone, bean dregs, bran-water and bean pulp as substrates by liquid state fermentation. The raw enzyme liquid was obtained by vacuum filtration after fermentation. The activity of hemieellulase of neurospora sitophila was determined with DNS method. The optimal fermentation medium, inoculating dosage and culture temperature for producing xylanase by hemicellulase was studied. The results showed that the best fermentation medium contained 3%bran-water, 0.5%peptone and 6%bean dregs, and the optimum inoculating dosage was 1%, and the process should be carried out under the conditions of natural pH at 28℃.