Shekhar Nilanshu, Bhattacharya Debaditya, Kumar Dishant, Gupta Rajinder K
School of Biotechnology, GGS Indraprastha University, Delhi, India.
Can J Microbiol. 2006 Sep;52(9):805-8. doi: 10.1139/w06-035.
During the previous decade, chitinases have received increased attention because of their wide range of applications. Chito-oligomers produced by enzymatic hydrolysis of chitin have been of interest in recent years because of their broad applications in medical, agricultural, and industrial applications, such as antibacterial, antifungal, hypo cholesterolemic, and antihypertensive activity, and as food quality enhancer. Fungal cell walls being rich in chitin also enable the use of chitinases in biocontrol of fungal pathogens, as bio-fungicides. An actinomycete was isolated from the bark of trees of Dehradun in India and was later identified as Streptomyces violaceusniger. This strain exhibits strong antagonism towards various wood-rotting fungi, such as Phanerochaete chrysosporium, Postia placenta, Coriolus versicolor, and Gloeophyllum trabeum. Further, studies showed an extracellular bioactive compound was responsible for the antagonism. The conditions for the production of this biocontrol agent were optimized, and the effects of various stress factors (like nitrogen-deficient media, carbon-deficient media, etc.) were studied. The presence of chitin in the growth media was found to be an essential factor for the active production of the biocontrol agent. The pH and temperature optima for the biocontrol agent were determined. Purification and characterization of this specific biocontrol agent was performed through anion exchange chromatography using a DEAE-cellulose column, and a single protein band was obtained on a 10% sodium dodecyl sulfate-polyacrylamide gel. The protein was later identified as a 28 kDa endo chitinase by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) and by a chitobiose activity assay.
在过去十年中,几丁质酶因其广泛的应用而受到越来越多的关注。近年来,几丁质经酶水解产生的壳寡糖因其在医学、农业和工业领域的广泛应用而备受关注,如抗菌、抗真菌、降胆固醇和降血压活性,以及作为食品品质增强剂。富含几丁质的真菌细胞壁也使得几丁质酶可作为生物杀真菌剂用于真菌病原体的生物防治。从印度德拉敦树木的树皮中分离出一种放线菌,后来鉴定为紫色链霉菌。该菌株对多种木材腐朽真菌表现出强烈的拮抗作用,如黄孢原毛平革菌、绒白耙齿菌、云芝和密粘褶菌。此外,研究表明一种细胞外生物活性化合物是造成这种拮抗作用的原因。优化了这种生物防治剂的生产条件,并研究了各种胁迫因素(如缺氮培养基、缺碳培养基等)的影响。发现生长培养基中几丁质的存在是生物防治剂活性产生的必要因素。确定了生物防治剂的最适pH值和温度。通过使用DEAE - 纤维素柱的阴离子交换色谱法对这种特定的生物防治剂进行了纯化和表征,在10%十二烷基硫酸钠 - 聚丙烯酰胺凝胶上获得了一条单一的蛋白带。该蛋白后来通过基质辅助激光解吸电离飞行时间质谱(MALDI - TOF)和壳二糖活性测定被鉴定为一种28 kDa的内切几丁质酶。