Cocoa pod husk, a new source of hydrolase enzymes for preparation of cross-linked enzyme aggregate

Abstract

Cocoa pod husk (CPH) is a by-product of cocoa production obtained after removing the beans from the fruit. The analysis of CPH has shown that it contains high amounts of protein. This study is aimed to utilize this protein source in hydrolase enzyme production. In this study, seven hydrolase enzymes (amylase, fructosyltransferase, mannanase, glucosidase, glucanase, lipase and protease) were screened from CPH for the first time for feasible industrial production. Among these hydrolases, lipase was chosen for the next steps of experiments as it has a lot of applications in different industries. The extraction of high active lipase from CPH has been done under optimum conditions. The condition that was optimum for the three major factors was achieved using Face centered central composite design (FCCCD) with response surface methodology (RSM) to obtain the highest enzyme activity of crude lipase from CPH. The optimum condition of extraction is used for preparation of cross-linked enzyme aggregate (CLEA). For the production of immobilized biocatalyst, the technique of CLEA is considered as an effective technique for its industrially attractive advantages. Referring to the results of OFAT, CLEA-lipase was prepared in the best condition at the presence of 30 mM ammonium sulphate, 70 mM glutaraldehyde with 0.23 mM Bovine serum albumin as an additive. Immobilization effectively improved the stability of lipase against various organic solvents.

Dual miRNases for Triple Incision of miRNA Target: Design Concept and Catalytic Performance

“…According to literature data, BSA is able to stabilize and enhance the activity of several natural enzymes including cellulose and restriction endonucleases, as well as commercially used proteases and lipases within cross-linked enzyme aggregates (CLEA) [50][51][52]. BSA possesses a high amount of positively charged amino acids lysines in its structure which defines the possible mechanisms of enzyme stabilization [51]. In particular, BSA may prevent the thermal denaturation of enzymes through direct binding and maintenance of their active conformation [52].…”

Determinação Do Fator De Efetividade Para Enzimas Imobilizadas Usando Métodos De Regressão Simbólica via Programação Genética

“…O processo de imobilização ocorre quando o glutaraldeído penetra na estrutura interna da enzima e seu grupo aldeído reage com os grupos amino da proteína que provoca uma agregação química entre as enzimas formando um "biocatalisador sólido", podendo eventualmente acontecer reações com outros grupos como os tióis, fenóis e imidazólios (YUSOF et al, 2016). Alguns fatores são importantes para controlar o tamanho final das partículas de CLEA, são eles: o tipo e quantidade do precipitado, a velocidade de agitação, concentração de proteínas, além da concentração de glutaraldeído (GARCIA-GALAN et al, 2011).…”

Enhanced Kinetic Performance of CLEA-Lipase Extracted from Skim Latex of Hevea brasiliensis upon Immobilization on Magnetic Iron Nanoparticles

“…It contains 75% of the total soluble proteins and some of these proteins are enzymes, including hydrolases [5]. According to Yusof et al [6], hydrolases are frequently used in the enzyme-based industries owing to the fact that, these enzymes do not require any co-factors, thus easy to use. Some of the industries that used enzymes are food manufacturing [7,8], detergents [9][10][11][12], pharmaceuticals [13][14][15] and biodiesel production [16][17][18][19].…”

“…Lipase, a type of hydrolase, has been purified from the latex serum of Hevea brasiliensis [20,21]. Furthermore, Yusof et al [5] had successfully immobilized this lipase via cross-linked enzyme aggregates (CLEA), a technique that immobilize enzymes without the need of supports [6,[22][23][24][25][26][27][28][29][30][31][32][33][34][35][36]. CLEA preparation involved precipitants, such as ammonium sulphate (NH4)2SO4, that can aggregate the enzymes, and polyfunctional cross-linker, such as glutaraldehyde, that links the enzyme aggregates together.…”