Banga J, Tripathi C K M
Fermentation Technology Division, Central Drug Research Institute, Lucknow, UP, India.
Lett Appl Microbiol. 2009 Aug;49(2):204-9. doi: 10.1111/j.1472-765X.2009.02640.x. Epub 2009 May 22.
Aim of the study was to develop a medium for optimal heparinase production with a strain of Aspergillus flavus (MTCC-8654) by using a multidimensional statistical approach.
Statistical optimization of intracellular heparinase production by A. flavus, a new isolate, was investigated. Plackett-Burman design was used to evaluate the affect of medium constituents on heparinase yield. The experimental results showed that the production of heparinase was dependent upon heparin, the inducer; chitin, structurally similar to heparin and NH(4)NO(3,) the nitrogen source. A central composite design was applied to derive a statistical model for optimizing the composition of the fermentation medium for the production of heparinase enzyme. The optimum fermentation medium consisted of (g l(-1)) Mannitol, 8.0; NH(4)NO(3), 2.5; K(2)HPO(4), 2.5; Na(2)HPO(4), 2.5; MgSO(4).7H(2)O, 0.5; Chitin, 17.1; Heparin, 0.6; trace salt solution (NaMoO(4).2H(2)O, CoCl(2).6H(2)O, CuSO(4).5H(2)O, FeSO(4).7H(2)O, CaCl(2)), 10(-4) mol l(-1).
A 2.37-fold increase in heparinase production was achieved in economic and effective manner by the application of statistical designs in medium optimization.
Heparinase production was doubled by statistical optimization in a cost-effective manner. This heparinase can find application in pharmaceutical industry and for the generation of low-molecular-weight heparins, active as antithrombotic and antitumour agents.
本研究的目的是通过多维统计方法，开发一种用于黄曲霉（MTCC - 8654）菌株最佳肝素酶生产的培养基。
对新分离的黄曲霉细胞内肝素酶生产进行了统计优化研究。采用Plackett - Burman设计评估培养基成分对肝素酶产量的影响。实验结果表明，肝素酶的产生依赖于诱导剂肝素、与肝素结构相似的几丁质以及氮源硝酸铵。应用中心复合设计得出统计模型，以优化用于肝素酶生产的发酵培养基组成。最佳发酵培养基由（g l⁻¹）甘露醇8.0、硝酸铵2.5、磷酸氢二钾2.5、磷酸氢二钠2.5、硫酸镁·7水合物0.5、几丁质17.1、肝素0.6、痕量盐溶液（钼酸钠·2水合物、氯化钴·6水合物、硫酸铜·5水合物、硫酸亚铁·7水合物、氯化钙）10⁻⁴ mol l⁻¹组成。
通过在培养基优化中应用统计设计，以经济有效的方式使肝素酶产量提高了2.37倍。
通过统计优化以具有成本效益的方式使肝素酶产量提高了一倍。这种肝素酶可应用于制药行业以及用于生产具有抗血栓和抗肿瘤活性的低分子量肝素。
