Purification and Characterization of an Extracellular Chitinase from the Antifungal Biocontrol Agent Streptomyces halstedii

2005 ◽  
Vol 27 (19) ◽  
pp. 1483-1486 ◽  
Author(s):  
Gil-Jae Joo
Keyword(s):  
Biocontrol Agent ◽  
Purification And Characterization ◽  
Streptomyces Halstedii

scholarly journals Purification and Characterization of Chitinase from the Nematode – Fungus Paecilomyces sp. P1

2019 ◽  
Vol 35 (1) ◽  
Author(s):  
Chu Thanh Binh ◽  
Nguyen Phuong Nhue ◽  
Ho Tuyen ◽  
Bui Thi Viet Ha
Keyword(s):  
Metarhizium Anisopliae ◽  
Biocontrol Agent ◽  
Specific Activity ◽  
Chitinase Activity ◽  
Paecilomyces Lilacinus ◽  
Purification And Characterization ◽  
Crinipellis Perniciosa ◽  
Isolation And Characterization ◽  
Invertebr Pathol

The nematophagous – fungi Paecilomyces sp. is curently developed as a biocontrol agent against plant parasitic nematodes (Khan et al., 2003; Yang et al., 2007). Biological control agents can infiltrate certain nematode sites and destroy them by producing some enzymes including chitinase (Khadijeh et al., 2017). The purpose of this study was to purify, determine the chitinase activity from Paecilomyces sp. P1. With Lugol reagent, chitinase of this strain was characterized by diffusion on agar plate. Chitinase specific activity was determined by measuring the release of reducing saccharides from colloidal chitin by the N-acetyl-glucosamine-dinitrosalicylate method at 540 nm. By using the saturated (NH4)2SO4 precipitation at 65% concentration, DEAE A-50 ion exchange chromatography and SDS - PAGE concentration 12.5%, chitinase molecules weigh nearly 50kDa, having a specific activity of 133,3 U/mg, 2,1-fold higher than that of supernatant. Furthermore, method of testing with the nematode Meloidogyne sp., the ability to kill nematodes of Paecilomyces sp. P1 reached 58% efficiency in 96h. These results were a scientific basis for the application of Paecilomyces sp. P1 in the production of nematode insecticides. Keywords Paecilomyces sp. P1; chitinase; purify, biocontrol, Meloidogyne sp References   [1] Nguyễn Ngọc Châu, Tuyến trùng thực vật và cơ sở phòng trừ, NXBKHKTHN, 2003.[2] Nguyễn Hữu Quân, Vũ Văn Hạnh, Quyền Đình Thi, Phạm Thị Huyền, Tinh sạch và đánh giá tính chất lý hóa của chitinase từ nấm Lecanicillium lecanii, Kỷ yếu Hội nghị Công nghệ Sinh học toàn quốc, 1 (2013) 426.[3] CM Baratto, V Dutra, JT Boldo, LB Leiria, MH Vainstein, A. Schrank Isolation, characterization and transcriptional analysis of the chitinase chi2 gene (DQ011663) from the biocontrol fungus Metarhizium anisopliae var. anisopliae., Curr Microbiol, 53 (2006) 217.[4] D. Wharton,. Nematode eggshells, Parasitology 81 (1980) 447.[5] F. A. Zaki, D. S. Bhatti , Effect of castor (Ricinus communus) and the biocontrol fungus Paecilomyces lilacinus on Meloidogyne javanica, Nematologica 36 (1980) 114.[6] H. M. Hussein Al Ajrami., Evaluation the Effect of Paecilomyces lilacinus as a Biocontrol Agent of Meloidogyne javanica on Tomato in Gaza Strip, Faculty of science Master of Biological Sciences Microbiology., 2016.[7] J. De la Cruz, A Hidalgo-Gallego, JM Lora, T Benitez, JA Pintor-Toro, A Llobell , Isolation and characterization of three chitinases from Trichoderma harzianum., Eur. J. Biochem,. 206 (1992) 859.[8] JLD Marco, MC Valadares-Inglis . Purification and characterization of an N-acetylglucosaminidase produced by a Trichodermaharzianum strain which controls Crinipellis perniciosa. Appl. Microbiol. Biotechnol. 64 (2003) 70.[9] JLD Marco , LHC Lima, MV Sousa MV, CR Felix A Trichoderma harzianum chitinase destroys the cell wall of the phytopathogen Crinipellis perniciosa, the causal agent of witches’ broomof cocoa, J Microbiol Biotechnol 16 (2000) 383.[10] Khan Alamgir, Williams Keith, Mark P. Molloy, and Nevalainen Henlena, Purification and characterization of a serine protease and chitinases from Paecilomyces lilacinus and detection of chitinase activity on 2D gels, Protein Expression and Purification 32 (2003) 210.[11] Khadijeh Abbsi, Doustmorad ZAFARI, Robert WICK., Evaluation of chitinase enzyme in fungal isolates obtained from golden potato cyst nematode (Globodera rostochiensis) Zemdirbyste-Agriculture, 2 (2017) 179.[12] Kopparapu Narasimha Kumar, Peng Zhou, Shuping Zhang, Qiaojuan Yan, Zhuqing Liu, Zhengqiang Jiang, Purification and characterization of a novel chitinase gene from Paecilomyces thermophila expressed in Escherichia coli. Carbonhydrate Reseach 347 (2012) 155.[13] Methanee Homthong, Anchanee Kubera, Matana Srihuttagum, Vipa Hongtrakul, Isolation and characterization of chitinase from soil fungi, Paecilomyces sp. Agriculture and Natural Resources, 1 (2016) 50.[14] RS Patil, V Ghormade, MV Desphande MV ,Chitinolytic enzymes: an exploration. Enzyme Microb. Technol. 26 (2002) 473[15] RJ Leger St , RM Cooper, AK Charnley, Characterization of chitinase and chitobiase produced by the entomopathogenic fungus Metarhizium anisopliae. J. Invertebr. Pathol. 58 (1991) 415.[16] S Leger, RJ Joshi RJ, RJ Bidochka, DW Roberts . Characterization and ultrastructural localization of Metarhizium anisopliae, M. xavoviride, and Beauveria bassiana during fungal invasion of host (Manduca sexta) cuticle. Appl Environ Microbiol 62 (1996)907.[17] SC Kang, S. Park, DG Lee ,, Purification and characterization of a novel chitinase from the entomopathogenic fungus, Metarhiziumanisopliae. J Invertebr Pathol., 73 (1999) 276.[18] P.J.M Bonants, P.F.L. Fitters, H. Thijs, E. den Belder, C. Waalwijk, J.W.D.M. Henfling. A basic serine protease from Paecilomyces lilacinus with biological activity against Meloidogyne hapla eggs, Microbiology 141(1995) 75.[19] VE Tikhonov, LV Lopez-Llorca, J Salinas, HB Jansson . Purification and characterization of chitinases from the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium, Fungal Genet Biol (2002) 67[20] Van Nam Nguyen, YJ Kim, KT Oh, WJ Jung, RD Park , The antifungal activity of chitinases from Trichoderma aureoviride DY-59 and Rhizopus microsporus VS-9. Curr. Microbiol 56 (2008) 28.[21] Van Nam Nguyen, In-Jae Oh, Young-Ju Kim, Kil-Yong Kim, Young-Cheol Kim, Ro-Dong Par J Ind., Purification and characterization of chitinases from Paecilomyces variotii DG-3 parasitizing on Meloidogyne incognita eggs, (2009) 195[22] Z. Perveen and S. Shahzad S., , A comparative study of the efficacy of Paecilomyces species against root-knot nematode Meloidogyne incognita. Pakistan Journal of Nematology, 31 (2013) 125

Cloning, expression, purification and characterization of HSP105

2010 ◽  
Vol 34 (8) ◽  
pp. S46-S46
Author(s):  
Dong Han ◽  
Huang Xu
Keyword(s):  
Purification And Characterization

Purification and characterization of phosphoglucomutase from peas

1994 ◽  
Vol 92 (3) ◽  
pp. 479-486 ◽  
Author(s):  
Cynthia M. Galloway ◽  
W. Mack Dugger
Keyword(s):  
Purification And Characterization

Purification and Characterization of Tissue-Type Plasminogen Activator in Maxillary Mucosa with Chronic Inflammation

1985 ◽  
Vol 54 (02) ◽  
pp. 485-489 ◽  
Author(s):  
Yukiyoshi Hamaguchi ◽  
Masuichi Ohi ◽  
Yasuo Sakakura ◽  
Yasuro Miyoshi
Keyword(s):  
Plasminogen Activator ◽  
Chronic Inflammation ◽  
Gel Filtration ◽  
Potassium Acetate ◽  
Tissue Type ◽  
Purification And Characterization ◽  
Tissue Type Plasminogen Activator ◽  
Urokinase Inhibitor ◽  
Type Plasminogen Activator

SummaryTissue-type plasminogen activator (TPA) was purified from maxillary mucosa with chronic inflammation and compared with urokinase. Purification procedure consisted of the extraction from delipidated mucosa with 0.3M potassium acetate buffer (pH 4.2), 66% saturation of ammonium sulfate, zinc chelate-Sepharose, concanavalin A-Sepharose and Sephadex G-100 gel filtration chromatographies.The molecular weight of the TPA was approximately 58,000 ± 3,000. Its activity was enhanced in the presence of fibrin and was quenched by placental urokinase inhibitor, but not quenched by anti-urokinase antibody. The TPA made no precipitin line against anti-urokinase antibody, while urokinase did.All these findings indicate that the TPA in maxillary mucosa with chronic inflammation is immunologically dissimilar to urokinase and in its affinity for fibrin.

Prothrombin Metz : Purification and Characterization of a Variant of Human Prothrombin

1979 ◽  
Author(s):  
M Ribieto ◽  
J Elion ◽  
D Labie ◽  
F Josso
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Factor Xa ◽  
Polyacrylamide Gel ◽  
Cleavage Pattern ◽  
Purification And Characterization ◽  
Double Immunodiffusion ◽  
The Family

For the purification of the abnormal prothrombin (Pt Metz), advantage has been taken of the existence in the family of three siblings who, being double heterozygotes for Pt Metz and a hypoprothrombinemia, have no normal Pt. Purification procedures included barium citrate adsorption and chromatography on DEAE Sephadex as for normal Pt. As opposed to some other variants (Pt Barcelona and Madrid), Pt Metz elutes as a single symetrical peak. By SDS polyacrylamide gel electrophoresis, this material is homogeneous and appears to have the same molecular weight as normal Pt. Comigration of normal and abnormal Pt in the absence of SDS, shows a double band suggesting an abnormal charge for the variant. Pt Metz exhibits an identity reaction with the control by double immunodiffusion. Upon activation by factor Xa, Pt Metz can generate amydolytic activity on Bz-Phe-Val-Arg-pNa (S2160), but only a very low clotting activity. Clear abnormalities are observed in the cleavage pattern of Pt Metz when monitored by SDS gel electrophoresis. The main feature are the accumulation of prethrombin l (Pl) and the appearance of abnormal intermediates migrating faster than Pl.

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