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Advances in Bioscience and Biotechnology 2014年第5期5卷 426-433页

作者： Tanzila Shahzadi Zahid Anwar Zafar Iqbal Awais Anjum Tahir Aqil Bakhtawar Arroha Afzal Muhammad Kamran Sajid Mehmood Muhammad Irshad Agriculture Biotechnology National Institute for Biotechnology and Genetic Engineering Faisalabad Pakistan Department of Biochemistry Nawaz Sharif Medical College University of Gujrat Gujrat Pakistan Department of Botany University of Gujrat Gujrat Pakistan Department of Microbiology Quaid-e-Azam University Islamabad Pakistan

In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth supporting substrate. Various physio-chemical and nutritional variables were optimized using classical and completely randomized designs for induced production of exoglucanase, and β-glucosidase from the co-culture of Trichoderma viride and Ganoderma lucidum in solid state fermentation (SSF). Analysis profile showed that when the conditions of the SSF medium containing 15 g corn stover substrate (50% w/w moisture) inoculated with 6 mL of inoculum were optimal, the maximum productions of exoglucanase (485 ± 6.5 U/mL) and β-glucosidase (255 ± 3.3 U/mL) were recorded after 5 days of incubation at pH 6 and 35°C.

关键词： Process Optimization SSF Co-Culture T. viride G. lucidum

Purification and characterization of the kinetic parameters of cellulase produced from wheat straw by Trichoderma viride under SSF and its detergent compatibility

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Advances in Bioscience and Biotechnology 2011年第3期2卷 149-156页

作者： Hafiz Muhammad Nasir Iqbal Ishtiaq Ahmed Muhammad Anjum Zia Muhammad Irfan Enzyme Biotechnology Laboratory Department of Chemistry and Biochemistry University of Agriculture Food Biotechnology Research Centre Pakistan Council of Scientific and Industrial Research Labs

This paper reports the purification and characterization of kinetic parameters of cellulase produced from Trichoderma viride under still culture solid state fermentation technique using cheap and an easily available agricultural waste material, wheat straw as growth supported substrate. Trichoderma viride was cultured in fermentation medium of wheat straw under some previously optimized growth conditions and maximum activity of 398±2.43U/mL obtained after stipulated fermentation time period. Cellulase was purified 2.33 fold with specific activity of 105U/mg in comparison to crude enzyme extract using ammonium sulfate precipitation, dialysis and Sephadex-G-100 column chromatography. The enzyme was shown to have a relative low molecular weight of 58kDa by sodium dodecyl sulphate poly-acrylamide gel electrophoresis. The purified enzyme displayed 6.5 and 55oC as an optimum pH and temperature respectively. Using carboxymethyl cellulose as substrate, the enzyme showed maximum activity (Vmax) of 148U/mL with its corresponding KM value of 68μM. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect on purified cellulase whereas, the enzyme activated by Co2+ and Mn2+ at a concentration of 1mM. The purified cellulase was compatible with four local detergent brands with up to 20 days of shelf life at room temperature suggesting its potential as a detergent additive for improved washing therefore, it is concluded that it may be potentially useful for industrial purposes especially for detergent and laundry industry.

关键词： Cellulase Trichoderma viride Purification SDS-PAGE Characterization Detergent Compatibility

A novel strain of Trichoderma virideshows complete lignocellulolytic activities

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Advances in Bioscience and Biotechnology 2012年第8期3卷 1160-1166页

作者： Kannan Neethu M. Rubeena S. Sajith S. Sreedevi Prakasan Priji K. N. Unni M. K. Sarath Josh V. N. Jisha S. Pradeep Sailas Benjamin Enzyme Technology Laboratory Biotechnology Division Department of Botany University of Calicut Malappuram India

In this study we describe a novel dark-green strain of Trichoderma viride exhibiting complete ensemble of cellulase, hemicellulase and ligninase activities on specific plate assays. To assess the cellulase production in detail, basal salt medium (BSM) was fortified with synthetic (carboxymethyl cellulose (CMC), glucose, sucrose, dextrose, lactose or maltose) and natural (flours of banana, banana peel, jack seed, potato or tapioca) carbon as well as nitrogen (yeast extract, beef extract, peptone, NaNO3 or NH4NO3) sources. Temperature and pH optima were 28°C and 4, respectively for the growth of the fungus in CMC-BSM with 137 U/mL cellulase activity, which was enhanced to 173 U/mL at 1.25% CMC concentration. Flours of potato and banana peel supported comparable yields of cellulase to that of CMC, while sodium nitrate was the preferred nitrogen source. The water soluble bluish-green pigment (a probable siderophore) extracted from the spores showed an absorption maximum at 292 nm. To sum up, the complete lignocellulolytic potential of this fungus offers great industrial significance, coupled with the production of a new pigment.

关键词： Trichoderma viride Morphology Liquid Fermentation Bluish-Green Pigment

Characterization of purified β-glucosidase produced from Trichoderma viridethrough bio-processing of orange peel waste

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Advances in Bioscience and Biotechnology 2013年第10期4卷 941-944页

作者： Muhammad Irshad Zahid Anwar Muhammad Ramzan Zahed Mahmood Haq Nawaz Department of Biochemistry NSMC University of Gujrat Gujrat Pakistan Department of Chemistry and Biochemistry University of Agriculture Faisalabad Pakistan Institute of Animal Nutrition & Feed Technology University of Agriculture Faisalabad Pakistan

In the present study, solid state fermentation was carried out using orange peel waste to produce β-glucosidase from Trichoderma viride. A locally isolated fungal strain T. viride was cultured in the solid state medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 515 ± 12.4 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Indigenously produced β-glucosidase was subjected to the ammonium sulfate precipitation and Sephadex-G-100 gel filtration chromatography. In comparison to the crude extract β-glucosidase was 5.1-fold purified with specific activity of 758 U/mg. The enzyme was shown to have a relative molecular weight of 62 kDa as evidenced by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The purified β-glucosidase displayed 6 and 60℃ as an optimum pH and temperature respectively.

关键词： Orange Peel Waste β-Glucosidase T. viride Purification SDS-PAGE Characterization

Establishment of HPLC Fingerprint, Cluster Analysis and Principle Component Analysis of Citri Reticulatae Pericarpium viride

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Medicinal Plant 2019年第1期10卷 69-73页

作者： Beibei JIN Xiangping PEI Huizhen LIANG School of Traditional Chinese Medicine Shanxi University of Chinese Medicine

[Objectives] This study aimed to establish HPLC fingerprint and conduct cluster analysis and principle component analysis for Citri Reticulatae Pericarpium viride. [Methods] Using the HPLC method, the determination was performed on XSelect~® HSS T3-C_(18) column with mobile phase of acetonitrile-0.5% acetic acid solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was 360 nm. The column temperature was 25℃. The sample size was 10 μL. With peak of hesperidin as the reference, HPLC fingerprints of 10 batches of Citri Reticulatae Pericarpium viride were determined. The similarity of the 10 batches of samples was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition) to determine the common peaks. Cluster analysis and principal component analysis were performed by using SPSS 17.0 statistical software. [Results] The HPLC fingerprints of the 10 batches of medicinal materials had total 11 common peaks, and the similarity was 0.919-1.000, indicating that the chemical composition of the 10 batches of medicinal materials was consistent. There were 11 common components in the 10 batches of medicinal materials, but their contents were different. When the Euclidean distance was 20, the 10 batches of samples were divided into two categories, S4 in the first category, and the others in the second one. When the Euclidean distance was 5, the second category could be further divided into two sub-categories, S1 and S10 in one sub-category, and S2, S3, S5, S6, S7, S8 and S9 in the other one. The principle component analysis showed that cumulative contribution rate of the two main component factors was 92.797%, and the comprehensive score of S7 was the highest with the best quality. [Conclusions] The results of HPLC fingerprinting, cluster analysis and principle component analysis can provide reference for the quality control of Citri Reticulatae Pericarpium viride.

关键词： Citri Reticulatae Pericarpium viride HPLC Fingerprint Cluster analysis Principle component analysis

Scab Disease Caused by Venturia inaequalison Apple Trees in Kyrgyzstan and Biological Agents to Control This Disease

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Advances in Microbiology 2017年第6期7卷 450-466页

作者： Tinatin Doolotkeldieva Saykal Bobusheva Plant Protection Department Faculty of Agriculture Kyrgyz-Turkish Manas University Bishkek Kyrgyzstan

Among the most economically important fruit crops in South and North Kyrgyzstan affected by scab disease are apples (Malus domestica). Natural isolates of Venturia inaequalis were isolated from diseased parts of apple trees, selected from different regions, through sedimentation of air microflora on the crowns of fruit trees. Two biocontrol agents as Trichoderma viride and Streptomyces sр. were tested for apple scab control in vitro and field conditions. Two applications of Trichoderma viride within 35 days completely stopped the scab disease in seedling leaves. Within 40 days, the seedlings were recovered;the new leaves have blossomed on branches. The Streptomyces sp. antagonists were inferior in effectiveness compared to Trichoderma viride;nevertheless, they prevented progression of the disease in the leaves. Seedlings grew new shoots and leaves around 50 days after the second treatment. Orchard experiment results provide great hope that a biological product based on Trichoderma viride could work as an effective agent to suppress the development of the pathogen in the early spring, when leaves start to show scab disease symptoms.

关键词： Apple Scab Venturia inaequalis Trichoderma viride Streptomyces sр. Biological Control

Enhanced bio-catalytic and tolerance properties of an indigenous cellulase through xerogel immobilization

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Advances in Bioscience and Biotechnology 2012年第4期3卷 308-313页

作者： Hafiz Muhammad Nasir Iqbal Shagufta Kamal Ishtiaq Ahmed Muhammad Tahir Naveed Department of Chemistry and Biochemistry University of Agriculture Faisalabad Pakistan Transgenic Lab Center of Agriculture Biochemistry and Biotechnology (CABB) University of Agriculture Faisalabad Pakistan

Today, demand exists for cost-effective production of industrially important enzymes from entire scientific sectors. By keeping in mind the extensive industrial applications of cellulase, this study was performed to immobilize the indigenous enzyme produced from Trichoderma viride under pre-optimized SSF of an agricultural waste material, wheat straw. To enhance the bio-catalytic and tolerance properties of the present enzyme gel matrix immobilization engineering was applied. Previously, 2.33~fold purified novel cellulase was immobilized in to a xerogel matrix of TMOS and PTMS. FTIR spectroscopy confirmed the successful immobilization of cellulase. The free and immobilized cellulase was characterized and stability profile showed that after 24 h incubation, immobilization enhanced the thermo-stability up to 75% against 80℃ as compare to the free enzyme. Xerogel matrix immobilization enhanced the catalytic efficiency of entrapped enzyme than that of the free cellulase. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect while, gel matrix immobilization enhanced 80% tolerance capacity of the cellulase against inactivating agents.

关键词： Bio-Catalysis Cellulase T. viride Immobilization Characterization

Characterization of Exo 1, 4-βglucanase produced from Tricoderma ViridiMBL through solid-state bio-processing of orange peel waste

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Advances in Bioscience and Biotechnology 2012年第5期3卷 580-584页

作者： Muhammad Irshad Zahid Anwar Amber Afroz Department of Biochemistry and Molecular Biology University of Gujrat Pakistan Department of Biochemistry NSMC University of Gujrat Pakistan Department of Chemistry and Biochemistry University of Agriculture Faisalabad Pakistan

Agro-industrial residues are primarily composed of complex polysaccharides that strengthen the microbial growth for the production of industrially important enzymes like cellulases. In the present study we aimed to characterize the Exo 1, 4-β glucanase that was indigenously produced from Trichoderma viride MBL. T. viride MBL was cultured in the Solid-State medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 412 ± 12 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Exo 1, 4-β glucanase was 4.17-fold purified with specific activity of 642 U/mg in comparison to the crude extract. To confirm its purity and molecular weight, sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS- PAGE) was performed. The enzyme was shown to have a molecular weight of 60 kDa with an optimum pH and temperature of 5 and 50℃, respectively. Lineweaver-Burk reciprocal plot revealed that the kinetic constants Km and Vmax of purified Exo 1, 4-β glucanase were 76 μM and 240 U/mL.

关键词： Orange Peel Waste Exo 1 4-β Glucanase T. viride MBL Purification SDS-PAGE

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三种生防木霉对五种烟田常用化合物的敏感性

植物保护学报 2015年第3期42卷 479-480页

作者：汪汉成王进黄艳飞王茂胜陆宁贵州省烟草科学研究院贵阳550081 长江大学生命科学学院湖北荆州434025 长江大学农学院湖北荆州434025

生防菌是植物病害防治中一类重要的生物资源,在土传病害(包括多种真菌和细菌病害)防治中起着重要作用(Pérez-Montaňo et al.,2014),其中有关木霉菌的报道较多(Hajieghrari et al.,2008;Schuster&Schmoll,2010),包括哈茨木霉Trichoderm... 详细信息

生防菌是植物病害防治中一类重要的生物资源,在土传病害(包括多种真菌和细菌病害)防治中起着重要作用(Pérez-Montaňo et al.,2014),其中有关木霉菌的报道较多(Hajieghrari et al.,2008;Schuster&Schmoll,2010),包括哈茨木霉Trichoderma harzianum、绿色木霉***和里氏木霉***-

关键词：木霉菌里氏木霉哈茨木霉绿色木霉土传病害细菌病害田常 viride 植物病害防治 Trichoderma

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测定L—赖氨酸的专用酶电极

氨基酸和生物资源 1985年第2期13卷 67-72页

作者： J.L.ROMETTE 孙泽辉汪庆松

自Trichoderma viride Y244—2的L—赖氨酸2—氧化酶固定于明胶载体后,固定在PO_2传感器上。所获酶电极用于连续流动系统,以便测定发酵罐内L—赖氨酸浓度。由于酶载体的性质所定,样品氧含量与信号的关系最小。酶电极的应答(response)确... 详细信息

自Trichoderma viride Y244—2的L—赖氨酸2—氧化酶固定于明胶载体后,固定在PO_2传感器上。所获酶电极用于连续流动系统,以便测定发酵罐内L—赖氨酸浓度。由于酶载体的性质所定,样品氧含量与信号的关系最小。酶电极的应答(response)确定在0.2mM到0.4mM赖氨酸浓度。用其他氨基酸试验了赖氨酸的特性。赖氨酸电极酶膜可用3000次贮存6个月稳定性良好。

关键词：酶电极酶膜明胶载体 viride 发酵罐葡萄糖氧化酶 Trichoderma 准曲线氧含量连续流动