Loschmidt Laboratories, Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment, Masaryk University, Brno, Czech Republic.
Biotechnol J. 2013 Jan;8(1):32-45. doi: 10.1002/biot.201100486. Epub 2012 Sep 11.
Haloalkane dehalogenases (EC 3.8.1.5, HLDs) are α/β-hydrolases which act to cleave carbon-halogen bonds. Due to their unique catalytic mechanism, broad substrate specificity and high robustness, the members of this enzyme family have been employed in several practical applications: (i) biocatalytic preparation of optically pure building-blocks for organic synthesis; (ii) recycling of by-products from chemical processes; (iii) bioremediation of toxic environmental pollutants; (iv) decontamination of warfare agents; (v) biosensing of environmental pollutants; and (vi) protein tagging for cell imaging and protein analysis. This review discusses the application of HLDs in the context of the biochemical properties of individual enzymes. Further extension of HLD uses within the field of biotechnology will require currently limiting factors - such as low expression, product inhibition, insufficient enzyme selectivity, low affinity and catalytic efficiency towards selected substrates, and instability in the presence of organic co-solvents - to be overcome. We propose that strategies based on protein engineering and isolation of novel HLDs from extremophilic microorganisms may offer solutions.
卤代烷烃脱卤酶(EC 3.8.1.5,HLD)是一种α/β-水解酶,可作用于裂解碳卤键。由于其独特的催化机制、广泛的底物特异性和高度的稳定性,该酶家族的成员已被应用于几种实际应用中:(i)生物催化制备有机合成中光学纯的构建块;(ii)化学过程副产物的回收;(iii)有毒环境污染物的生物修复;(iv)战争制剂的净化;(v)环境污染物的生物传感;以及(vi)细胞成像和蛋白质分析的蛋白质标记。本综述讨论了 HLD 在个别酶的生化特性方面的应用。要在生物技术领域进一步扩展 HLD 的用途,就需要克服目前的限制因素,如低表达、产物抑制、酶选择性不足、对选定底物的低亲和力和催化效率、以及在有机溶剂共溶剂存在下的不稳定性。我们提出,基于蛋白质工程和从极端微生物中分离新型 HLD 的策略可能提供解决方案。