Ramya L N, Pulicherla K K
Department of Biotechnology, Acharya Nagarjuna University, Guntur, 522510 A.P. India.
Center for Bioseparation Technology, VIT University, Vellore, 632014 TN India.
J Food Sci Technol. 2015 Sep;52(9):5484-96. doi: 10.1007/s13197-014-1654-6. Epub 2014 Nov 26.
Pectin is a complex structural heteropolysaccharide that require numerous pectinolytic enzymes for its complete degradation. Polygalacturonase from mesophilic or thermophilic origin are being widely used in fruit and vegetable processing in the recent decades to degrade pectic substances. Recently cold active pectinases are finding added advantages over meso and thermophilic counterparts, to use in industrial scale particularly in food processing industry. They facilitate in conservation of several properties of foods so that the end product retains its naturality and also generates economic benefits. In the present study, Pseudoalteromonas haloplanktis, a well reported marine psychrophile is taken as a model organism for cold active polygalacturonase and is evaluated in comparision to the routinely used mesophilic and thermophilic enzymes by insicio approach. Polygalacturonase sequences from industrially important microbial sources were subjected to MEME and Pfam wherein motifs and domains involved in the conservation were analyzed. Dendrogram revealed sequence level similarity and motifs showed uniform distribution of conserved regions that are involved in important functions. It was also observed through clustalW analysis that the amount of arginine content of psychrophiles is less when compared with thermophiles. Finally, all the modeled enzyme structures were subjected to docking studies using Autodock 4.2 with the substrate polygalacturonic acid and binding energies were found to be -5.73, -6.22 and -7.27 KCals/mole for meso, thermo and psychrophiles respectively which indicates the efficiency of psychrophilic enzymes when compared with its counterparts giving scope for further experimentation to find their better usage in various food industry applications.
果胶是一种复杂的结构性杂多糖,其完全降解需要多种果胶分解酶。近几十年来,来自嗜温或嗜热菌的聚半乳糖醛酸酶在果蔬加工中被广泛用于降解果胶物质。最近,冷活性果胶酶相对于中温及嗜热果胶酶显示出额外的优势,尤其在食品加工业等工业规模应用中。它们有助于保留食品的多种特性,使最终产品保持天然性并产生经济效益。在本研究中,假交替单胞菌属嗜冷海洋菌被用作冷活性聚半乳糖醛酸酶的模式生物,并通过原位方法与常规使用的中温及嗜热酶进行比较评估。对来自具有工业重要性的微生物来源的聚半乳糖醛酸酶序列进行了MEME和Pfam分析,其中对参与保守性的基序和结构域进行了分析。系统发育树揭示了序列水平的相似性,基序显示了参与重要功能的保守区域的均匀分布。通过ClustalW分析还观察到,嗜冷菌的精氨酸含量与嗜热菌相比更少。最后,所有建模的酶结构都使用Autodock 4.2与底物聚半乳糖醛酸进行对接研究,发现中温、嗜热和嗜冷菌的结合能分别为-5.73、-6.22和-7.27千卡/摩尔,这表明嗜冷酶与其同类酶相比具有更高的效率,为进一步实验以探索其在各种食品工业应用中的更好用途提供了空间。