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.
Highlights
Cocoa pod husk (CPH) is a waste by-product of the cocoa industry, obtained after the removal of the cocoa beans from the fruit
Stability of cross-linked enzyme aggregate (CLEA)‐lipase against hydrophilic organic solvents In our previous work, the CLEA-lipase from CPH was characterized and the results indicated that the immobilized lipase has superior stability in response to different ranges of temperature (25–60 °C) and pH (5–10) in comparison to free form of the enzyme (Khanahmadi et al 2015)
The enzyme molecules are packed in a small volume, where mass-transport limitations will surely be expected, especially with fast reactions. This effect is found to be small when the CLEA is dispersed in the solution at the end of the cross-linking process. In this project, seven of the most important hydrolase enzymes from an agro waste byproduct, CPH have been screened for the first time successfully
Summary
Cocoa pod husk (CPH) is a waste by-product of the cocoa industry, obtained after the removal of the cocoa beans from the fruit. 52–76 % of the weight of the cocoa fruit is pod husk (Donkoh et al 1991). For each ton of dry beans produced, ten tons of cocoa pod husks are generated, which presents a serious challenge for waste management (Figuiera et al 1993). It may be a significant source of disease inocula, such as black pod rot (Figuiera et al 1993; Barazarte et al 2008). In 90s, some researchers chemical analysis of CPH showed high percentage of crude protein, between 70 and 90 g/kg (Donkoh et al 1991; Vriesmann et al 2012). Since it is recognised that, extraction of the protein from this CPH has immense economic advantages for the cocoa producer countries and it can reduce some of the environmental problems as well
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