利用羧甲基纤维钠(CMCNa)平板筛选法,从白酒发酵副产物黄水中分离得到12株产纤维素酶菌株,其中菌株M34和菌株N2的比酶活最大,被选为后续研究对象。根据细菌形态特征观察,生理生化特性分析并结合16S r RNA序列分析,鉴定菌株M34和菌株N2分别为环状芽孢杆菌(Bacillus circulans)和内生芽孢杆菌(Bacillus endophyticus)。菌株经液态发酵培养,运用DNS法测定纤维素酶系酶活力,结果表明菌株N2的各酶活均高于菌株M34,其羧甲基纤维素酶活为0.132 U/m L,微晶纤维素酶活为0.012 U/m L,滤纸酶活为0.041 U/m L,β-葡萄糖苷酶活为0.158 U/m L。

12 bacteria w ith cellulase-producing capacity w ere isolated from the by-productof H uangshui(ferm ented m ash)from C hinese liquorfer-m entation by carboxym ethylcellulose(C M C)plate screening m ethod.Tw o strainsnam ed M 34 and N 2 w ith high specific enzym e activity w ere chosen asthe follow -up testobject.A ccording to theirm orphology features,biochem icaland physiologicalcharacteristics and 16S rR N A sequence analysis, strain M 34 w as identified as B acillus circulans and strain N 2 w as identified as B acillus endophyticus.B oth strains cellulase activities w ere detected by D N S m ethod and theresultsshow ed thattheenzym eactivity ofstrain N 2 w ashigherthan M 34.A m ong the enzym e activity ofstrain N 2,the C M C ase activity w as0.132 U /m l,the avicellulase activity w as0.012 U /m l,the FPase activity w as0.041 U /m land the β-glucosidase activity w as0.158 U /m l.

应用低能氮离子(N+)注入技术对纤维素酶产生菌里氏木霉(Trichoderma reesei)进行诱变选育,在能量为10 keV,注量为150×1014和200×1014N+/cm2的条件下分别筛选得到3株纤维素酶高产菌株,连续5代遗传稳定性实验结果表明,所得到的高产菌株遗传稳定性较好,羧甲基纤维素酶活力均提高到3.300 IU/mL以上,较出发菌株(2.698 IU/mL)提高了20.0%以上。采用Plackett-Burman实验设计法和旋转中心组合设计法系统地研究高产菌株150-1-1发酵营养因子组成,得到了纤维素酶产量随葡萄糖、麸皮和微晶纤维素等营养因子的变化规律及相应的响应面分析图。实验结果表明,葡萄糖、麸皮和微晶纤维素浓度与纤维素酶活存在显著的相关性,当葡萄糖浓度为4.9 g/L,麸皮浓度为23.0 g/L,微晶纤维素浓度为7.7 g/L时,150-1-1纤维素酶滤纸酶活力达到2.439 IU/mL,较优化前(2.000 IU/mL)提高了22.0%。

Cellulase producing strain Trichoderma reesei was mutated by low energy N + implantation.When the implantation energy was 10 keV,and the implantation doses were 150 ×1014 and 200 ×1014 N +/cm2,three high yield mutant strains were obtained.Their cellulase yield reached 3.300 IU/mL,and improved over 20.0% than that of original strain(2.698 IU/mL). Furthermore,mutant strains had better heredity stability after five passages.Plackett-Burman and central composite rotatable design experiment were applied to optimize the concentration of nutrient factors for cellulase production.The changing patterns of glucose,wheat bran and microcrystalline cellulose were obtained,and the corresponding response surface analysis diagram were also obtained.Experimental results showed that glucose,wheat bran and microcrystalline cellulose had an individual significant influence on cellulose production.The optimum concentration of nutrient factors for cellulase production was 4.9 g/L of glucose, 23.

以羧甲基纤维素酶活力、滤纸酶活力为考察指标，探讨绿色木霉（Trichoderma viride ）化L4C 和黑曲霉（Aspergillus niger ）混合固态发酵产纤维素酶的最佳培养时间；并以降解率为考察指标，通过单因素和正交试验对混菌固态发酵产纤维素酶条件进行优化，以促进秸秆资源的饲料化利用、缓解粮食危机，降低环境污染。结果表明，绿色木霉化 L4C 和黑曲霉混合固态发酵最佳时间为3 d，最佳总接种量为10％，绿色木霉化 L4C 与黑曲霉的最佳接种比例为1∶1，培养基最佳氮源为硫酸铵，麸皮与稻草秸秆粉最佳质量比为3∶7，最适宜培养基含水量为50％。在该条件下稻草秸秆半纤维素降解率可达34.83％，纤维素降解率可达39.75％，木质素降解率可达27.41％。

The optimum fermentation time of solid fermentation of mixed Trichoderma viride L4C and Aspergillus niger with filter paper activity and carboxymethyl cellulase activity as indexes was studied,and the solid fermentation conditions were optimized by single factor test and orthogonal experiments with degradation rate as index,so as to promote the utilization of straw for fodder,relieving food crisis,reducing the environmental pollution.The optimum solid fermentation conditions were fermentation time of 3 d,total inoculum volume of 10%,inoculation ratio of Trichoderma viride L4C to Aspergillus niger of 1 ∶ 1,(NH4 )2 SO4 as nitrogen source, mass ratio of bran to straw stalk powder of 3 ∶ 7,moisture contents of 50%.Under the above conditions,the degradation rate of hemicellulose,cellulose,lignin for straw stalk were 34·83%, 39·75% and 27·41%.

以不同植物中分离到的4株内生球毛壳菌NK102、NK103、NK104和NK105为对象,研究不同生态来源球毛壳菌降解木质素和纤维素的能力。方法:首先采用羧甲基纤维素和纤维素刚果红平板检测各菌株的纤维素降解能力,并利用Bavendamm平板反应检测各菌株的木质素降解能力;将4株菌分别培养在以微晶纤维素、杨树叶和木屑为惟一碳源的液体培养基中,通过检测培养液中纤维素酶和漆酶的酶活力,比较各菌株分解利用天然木质纤维素材料的能力,连续培养12 d后检测培养液中次级代谢产物的合成情况;利用已测序的球毛壳菌CBS148.51的基因组信息,寻找编码木质纤维素降解酶类的基因,为球毛壳菌分解利用木质纤维素提供分子生物学依据。结果:NK102、NK103、NK104和NK105在羧甲基纤维素培养基和纤维素刚果红培养基上都能够生长并形成水解圈;Bavendamm平板反应显示4株菌降解木质素的能力由强到弱依次是NK103、NK102、NK105和NK104。4株菌都能分解利用微晶纤维素、杨树叶和木屑,分泌纤维素酶和漆酶,其中NK102在以木屑为碳源的培养基上纤维素酶活力最强,达到0.76 U/mL发酵液,NK103在以杨树叶为碳源的培养基上漆酶活力最强。与此同时,4株菌在发酵培养过程中都能够稳定地合成球毛壳甲素(ChA),ChA产量受到碳源影响,在以杨树叶为碳源的培养基上,NK104的ChA产量最高,可达到14.88 mg/L发酵液。利用已测序的球毛壳菌CBS148.51的基因组信息,寻找到119个编码纤维素半纤维素酶的基因、8个编码漆酶的基因和2个编码锰过氧化物酶的基因,球毛壳菌具有完整的降解纤维素半纤维素的酶体系,在木质纤维素降解真菌的开发过程中具有重要的研究价值。结论:本研究为球毛壳菌木质纤维素降解过程的研究及该菌种的开发利用奠定了基础。

Objective: The decomposition abilities of lignocellulosic materials by Chaetomium globosum NK102, NK103, NK104 and NK105, endophytes from different plants were evaluated. Methods: The cellulose utilizing ca-pability of the C.globosum isolates were tested on carboxymethylcellulose agar and cellulose-congo red agar. The lignin utilizing experiments were performed on Bavendamm plates, and the degradative activity was compared by measuring the respective zone of color change. The lignocellulases production potential of these isolates using mi-crocrystalline cellulose, the leaves of Populus sp. and wood powder as the sole carbon source in liquid fermenta-tion was assessed. In addition, secondary metabolites produced by the C.globosum isolates were detected after 12 days cultivation. In the sequenced genome of C.globosum CBS148.51, genes encoding enzymes involved in lignocel-luloses degrading were identified by sequence homology alignment. Results: C.globosum NK102, NK103, NK104 and NK105 fo

目的：观察虎杖水煎液对D-半乳糖（D-g a l）所致衰老小鼠学习、记忆功能的影响，并初步探讨其作用机制。方法取I C R小鼠80只，雌雄兼用，随机分成4组：正常对照组（NG，n=20），模型对照组（MG，n=20），虎杖水煎液低剂量组（LG，5g/kg，n=20），虎杖水煎液高剂量组（HG，10g/kg， n=20）。正常对照组给予背部皮下注射生理盐水，其余各组用 D-gal按150mg/kg背部皮下注射6周，每日一次，造成亚急性衰老模型。造模同时，虎杖水煎液低剂量组和高剂量组分别给予相应浓度的虎杖水煎液，正常对照组和模型对照组给予相同容积的0.5%羧甲基纤维素钠（CMC）。采用Y型电迷宫检测学习、记忆功能；使用相关试剂盒测定脑皮层中超氧化物歧化酶（SOD）和单胺氧化酶-B（MAO-B）的活力及丙二醛（MDA）和脂褐质（LP）的含量。结果给予D-gal 6周后，Y迷宫检测模型组记忆功能明显下降，表现为不同时间的正确反应率降低，潜伏期延长；而虎杖水煎液能明显缩短衰老小鼠潜伏期，显著升高正确反应率。生化检测指标显示，模型组脑皮层中MDA和LP含量升高；虎杖水煎液能降低脑皮层中MAO-B活性，升高SOD活性，降低脑皮层中MDA和LP含量。结论虎杖水煎液对D-gal诱导衰老模型小鼠的记忆功能有保护作用，其机制可能与其减少脑中MDA及LP含量，降低MAO-B活性，升高SOD活性有关。

Aim: To investigate the effect of Rhizoma Polygoni Cuspidati (RPC) decoction on learning and memory function of aging model mice induced by D-galactose, and explore its mechanism preliminarily. Methods: 80 ICR mice, male and female were both used, were randomly divided into 4 groups: Normal control group (NG, n=20), Model control group (MG, n=20), Low-dose drug treatment group (LG,5g/kg,n=20), High-dose drug treatment group (HG,10g/kg,n=20).Except NG was subeutaneously injected with saline, other groups were given subcutaneously injection of D-galactose (D-gal) at 150 mg/kg once a day for six weeks to induce subacute aging model. At the same time LG was oral y administrated RPC decoction at dose 5g/kg and HG at 10g/kg. NG and MG were given oral y the same volume 0.5% carboxymethyl cellulose sodium(CMC). Y-maze was use to determine the function of learning and memory. The analytic kit was used to measure the content of malondialdehyde (MDA) and lipofuscin (LP), the activity of superoxid