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Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification

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 hig...

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Main Authors: Sheladiya, Priyanka, Kapadia, Chintan, Prajapati, Vimal, El Enshasy, Hesham Ali, Abd. Malek, Roslinda, Marraiki, Najat, Zaghloul, Nouf S. S., Sayyed, R. Z.
Format: Article
Language:English
Published: Nature Research 2022
Subjects:
Q Science (General)
Online Access:http://eprints.utm.my/103977/1/HeshamAli2022_ProductionStatisticalOptimizationandFunctional.pdf
http://eprints.utm.my/103977/
http://dx.doi.org/10.1038/s41598-022-11022-0
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spelling my.utm.1039772023-12-11T01:49:43Z http://eprints.utm.my/103977/ Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification Sheladiya, Priyanka Kapadia, Chintan Prajapati, Vimal El Enshasy, Hesham Ali Abd. Malek, Roslinda Marraiki, Najat Zaghloul, Nouf S. S. Sayyed, R. Z. Q Science (General) 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. Nature Research 2022-12 Article PeerReviewed application/pdf en http://eprints.utm.my/103977/1/HeshamAli2022_ProductionStatisticalOptimizationandFunctional.pdf Sheladiya, Priyanka and Kapadia, Chintan and Prajapati, Vimal and El Enshasy, Hesham Ali and Abd. Malek, Roslinda and Marraiki, Najat and Zaghloul, Nouf S. S. and Sayyed, R. Z. (2022) Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification. Scientific Reports, 12 (1). pp. 1-16. ISSN 2045-2322 http://dx.doi.org/10.1038/s41598-022-11022-0 DOI:10.1038/s41598-022-11022-0
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
language English
topic Q Science (General)
spellingShingle Q Science (General)
Sheladiya, Priyanka
Kapadia, Chintan
Prajapati, Vimal
El Enshasy, Hesham Ali
Abd. Malek, Roslinda
Marraiki, Najat
Zaghloul, Nouf S. S.
Sayyed, R. Z.
Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification
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.
format Article
author Sheladiya, Priyanka
Kapadia, Chintan
Prajapati, Vimal
El Enshasy, Hesham Ali
Abd. Malek, Roslinda
Marraiki, Najat
Zaghloul, Nouf S. S.
Sayyed, R. Z.
author_facet Sheladiya, Priyanka
Kapadia, Chintan
Prajapati, Vimal
El Enshasy, Hesham Ali
Abd. Malek, Roslinda
Marraiki, Najat
Zaghloul, Nouf S. S.
Sayyed, R. Z.
author_sort Sheladiya, Priyanka
title Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification
title_short Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification
title_full Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification
title_fullStr Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification
title_full_unstemmed Production, statistical optimization, and functional characterization of alkali stable pectate lyase of Paenibacillus lactis PKC5 for use in juice clarification
title_sort production, statistical optimization, and functional characterization of alkali stable pectate lyase of paenibacillus lactis pkc5 for use in juice clarification
publisher Nature Research
publishDate 2022
url http://eprints.utm.my/103977/1/HeshamAli2022_ProductionStatisticalOptimizationandFunctional.pdf
http://eprints.utm.my/103977/
http://dx.doi.org/10.1038/s41598-022-11022-0
_version_ 1787132167812284416
score 13.211869

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