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Purification and Some Properties of Endo-1,4-β-glucanases of Trichoderma harzianum UzCF-28

Open Journal of Applied Sciences 2016年第8期6卷 514-523页

作者： N. Sh. Azimova D. M. Khamidov M. B. Djumagulov Z. S. Shakirov Institute of Microbiology of Uzbekistan Academy of Sciences Tashkent Republic of Uzbekistan

This work aimed at isolation, purification and study of biochemical features of cellulolytic enzymes synthesized by Trichoderma harzianum UzCF-28 strain. Strain UzCF-28 revealed a high cellulolytic activity during submerged cultivation in the liquid culture on modified Mandels nutrient medium, where wheat straw was used as a source of carbon. As a result of purification by precipitation with ammonium sulfate and further ion exchange chromatography, two isoforms of endo- 1,4-β-glucanase-EG II and EG III with molecular weight of 135 and 75 kDa respectively were revealed. The pH optimum for EG I and EG III was 4.5, while for EG II—4.7, irrespective of the applied substrates—either CMC or “Whatman filter” paper. Heating up to 40°C of EG III did not lead to its inactivation, and on the contrary, its activity increased by more than three times comparing to the initial activity of the enzyme, i.e. thermostability of EG III among tested enzymes significantly varied.

关键词： Trichoderma harzianum Enzyme Endo-1 4-β-glucanase Purification

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PtrCel9A7,an auxin-induced EGase in poplar,is involved in cell expansion and lateral root formation

PtrCel9A7,an auxin-induced EGase in poplar,is involved in ce...

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2017中国长三角遗传学大会

作者： Liangliang Yu Laigeng Li Shanghai Key Lab of Bio-energy Crops School of Life SciencesShanghai University National Key Laboratory of Plant Molecular Genetics/CAS Key Laboratory of Synthetic Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of Sciences

Plant endo-1,4-β-glucanases(EGases,EC 3.***.1.4),often referred to as cellulases,play important roles in many physiological processes in plant growth and development,such as cell expansion,fruit ripening,and organ *** to phylogenetical relationship,plant EGase gene family can be grouped into α,β and γ subfamilies[1].In our previous study,we identified a xylem-specific β-type EGase,PtrCel9 A6,in Populus,and characterized its roles in hydrolyzing amorphous cellulose and secondary cell wall formation in developing xylem[2].PtrCel9 A7,another β-type EGase member in Populus,is a close relative of PtrCel9 A6,but shows a different expression profile from *** qRT-PCR and promoter-GUS reporter systems,we found that PtrCel9 A7 is expressed in leaves,stems,flowers,shoot tips,and *** roots,GUS staining was specifically detected in lateral root(LR) primodia and basal part of LRs,indicating a potential function of PtrCel9 A7 in LR *** auxinresponsive elements(TGTCTC) were found in the promoter region of PtrCel9 A7,and treatment of 10μM IAA strongly induced expression of PtrCel9 A7 in the PtrCel9 A7 pro:GUS transgenic root system,while the induction was not observed with treatments of other *** expression of PtrCel9 A7 driven by CaMV 35 S promoter in Arabidopsis enhanced plant growth by increasing cell size,consistent with the results of PtrCel9 A6 overexpression[2],indicating a conserved function of R-type EGases in regulating cell ***,overexpression of PtrCel9 A7 significantly increased LR number,demonstrating a novel role of PtrCel9 A7 in LR *** that auxin plays a pivotal role during LR initiation and emergence and induces PtrCel9 A7 expression in roots,we speculate that PtrCel9 A7 acts downstream of auxin signaling to regulate LR formation.

关键词： endo-l 4-β-glucanase cell expansion lateral root Populus

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