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Computational modelling approach for the optimization of apple juice clarification using immobilized pectinase and xylanase enzymes.

ABSTRACT: Apple juice is typically marketed as a clear juice, and hence enzymatic treatments are common practices in juice industry. However, enzymatic treatments have been shown to face some challenges when process efficiency, and cost effectiveness are concerned. Therefore, it is necessary to optimize the enzymatic treatment process to maximize efficiency, and reuse enzymes to minimize the overall cost via immobilization. In this context, the present work features the immobilization of pectinase and xylanase from M. hiemalis on genipin-activated alginate beads, with subsequent evaluation of its efficacy in apple juice clarification. A central composite rotatable design (CCRD), coupled with artificial neural network (ANN) for modeling and optimization was used to design the experiments. Deploying a coupling time up to 120 min, and an agitation rate of 213 rpm (pectinase) - 250 rpm (xylanase), a maximum fractional enzyme activity recovered was observed to be about 81-83% for both enzymes. Optimum enzyme loading and genipin concentration were found to be 50 U/ml and 12% (w/v), respectively. The immobilized enzyme preparations were suitable for up to 5 repeated process cycles, losing about 45% (pectinase) - 49% (xylanase) of their initial activity during this time. The maximum clarity of apple juice (%T₆₆₀, 84%) was achieved at 100 min when pectinase (50 U/ml of juice) and xylanase (20 U/ml of juice) were used in combination at 57 °C. The immobilized enzymes are of industrial relevance in terms of biocompatibility, recoverability, and operational-storage stability.

SUBMITTER: Hassan SS

PROVIDER: S-EPMC7680705 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Computational modelling approach for the optimization of apple juice clarification using immobilized pectinase and xylanase enzymes.

Hassan Shady S SS Williams Gwilym A GA Jaiswal Amit K AK

Current research in food science 20201014

Apple juice is typically marketed as a clear juice, and hence enzymatic treatments are common practices in juice industry. However, enzymatic treatments have been shown to face some challenges when process efficiency, and cost effectiveness are concerned. Therefore, it is necessary to optimize the enzymatic treatment process to maximize efficiency, and reuse enzymes to minimize the overall cost via immobilization. In this context, the present work features the immobilization of pectinase and xyl ...[more]

PMID: 33251526

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Project description:BackgroundThe extracellular xylanase secreted by microorganisms is a hydrolytic enzyme, which arbitrarily cleaves the β-1, 4 backbone of the polysaccharide xylan; an enzyme used in the food processing, bio-pulping and bio-bleaching. The commercial production of the xylanase is limited because of a higher cost involvement, which can be overcome by the cost-effective production of the xylanase through immobilization of the microbial cell by the non-toxic substances.ObjectivesIn this work, the optimization of the extra-cellular cellulase free xylanase production by the immobilized cell of the Bacillus pumilus IMAU80221 strain using Ca-alginate beads along with standardization of the various parameters for a higher xylanase production were studied.Materials and methodsFollowing to sterilization, the Na-alginate solution was mixed with the bacterial suspension of the Bacillus pumilus IMAU80221 and was added drop by drop into the 1 M calcium chloride solution for 1 h for obtaining a uniform sized polymeric bead of the Ca-alginate. For xylanase production, the Ca-alginate beads were then transferred into 100 mL Erlenmeyer flasks with 20 mL of the culture medium containing (w/v) 0.02% NaCl, 0.02% MgSO4, 0.04% (KH4)2PO4, 0.1% peptone, and 0.5% xylan and incubated at 34 °C in an incubator shaker (150 rpm) for 24 h. The resultant supernatant (crude enzyme) was used for enzyme assay.ResultsThe maximum xylanase production by the free cell (1.9 U.mL-1.min-1) was recorded at 48 h which was 40.5% lower than the xylanase production by the immobilized cell (2.67 U.mL-1.min-1) at the same time. The beads containing the immobilized cells could be reused up to eight fermentation cycles for xylanase production and retained 83.5% of the productivity at the fourth cycle. The entrapped cells were stable after six months of storage at 4 °C and retained 68% of the xylanase productivity.ConclusionCellulase free xylanase production from the immobilized Bacillus pumilus IMAU80221 was optimized. The xylanase production by the immobilized cells of Bacillus pumilus was higher by 40.5 and 132.6 % over the free cells respectively after 48 and 72 h of the incubation.

Project description:Pectate lyase is a hydrolytic enzyme used by diverse industries to clarify food. The enzyme occupies a 25% share of the total enzyme used in food industries, and their demand is increasing gradually. Most of the enzymes in the market belong to the fungal origin and take more time to produce with high viscosity in the fermentation medium, limiting its use. The bacteria belonging to the genus Bacillus have vast potential to produce diverse metabolites of industrial importance. The present experiment aimed to isolate pectate lyase-producing bacteria that can tolerate an alkaline environment at moderate temperatures. Bacillus subtilis PKC2, Bacillus licheniformis PKC4, Paenibacillus lactis PKC5, and Bacillus sonorensis ADCN produced pectate lyase. The Paenibacillus lactis PKC5 gave the highest protein at 48 h of incubation that was partially purified using 80% acetone and ammonium sulphate. Purification with 80% acetone resulted in a good enzyme yield with higher activity. SDS-PAGE revealed the presence of 44 kDa molecular weight of purified enzyme. The purified enzyme exhibits stability at diverse temperature and pH ranges, the maximum at 50 °C and 8.0 pH. The metal ions such as Mg2+, Zn2+, Fe2+, and Co2+ significantly positively affect enzyme activity, while increasing the metal ion concentration to 5 mM showed detrimental effects on the enzyme activity. The organic solvents such as methanol and chloroform at 25% final concentration improved the enzyme activity. On the other hand, detergent showed inhibitory effects at 0.05% and 1% concentration. Pectate lyase from Paenibacillus lactis PKC5 had Km and Vmax values as 8.90 mg/ml and 4.578 μmol/ml/min. The Plackett-Burman and CCD designs were used to identify the significant process parameters, and optimum concentrations were found to be pectin (5 gm%) and ammonium sulphate (0.3 gm%). During incubation with pectate lyase, the clarity percentage of the grape juice, apple juice, and orange juice was 60.37%, 59.36%, and 49.91%, respectively.

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Computational modelling approach for the optimization of apple juice clarification using immobilized pectinase and xylanase enzymes.

Publications

Computational modelling approach for the optimization of apple juice clarification using immobilized pectinase and xylanase enzymes.

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