Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics

Starch hydrolyzing amylases are among the vital enzymes used in the food, textile, detergent, and pharmaceutical industries. The aim was to isolate amylase bacteria from agricultural soil, its extraction and subsequent purification. Within the scope of this work, using a 1% starch agar medium, poten...

Full description

Saved in:
Bibliographic Details
Main Authors: Ume-e-Salma Liaqat, Shumaila Naz, Hafiz Iftikhar Hussain
Format: Article
Language:English
Published: Elsevier 2024-01-01
Series:Kuwait Journal of Science
Subjects:
Online Access:https://www.sciencedirect.com/science/article/pii/S2307410823001979
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849735889790435328
author Ume-e-Salma Liaqat
Shumaila Naz
Hafiz Iftikhar Hussain
author_facet Ume-e-Salma Liaqat
Shumaila Naz
Hafiz Iftikhar Hussain
author_sort Ume-e-Salma Liaqat
collection DOAJ
description Starch hydrolyzing amylases are among the vital enzymes used in the food, textile, detergent, and pharmaceutical industries. The aim was to isolate amylase bacteria from agricultural soil, its extraction and subsequent purification. Within the scope of this work, using a 1% starch agar medium, potent thermostable bacterial amylase (A-4) was isolated from agricultural soil samples. Molecular analysis revealed the amylase-producing bacterial strain (A-4) to be Citrobacter portucalensis. We used agro-industrial-based wastes (wheat bran, potato peel, and sugarcane bagasse) in amylase production. Culture condition optimization showed that high amylase activity of 4.96 U/mL was obtained at temperature 55 °C and pH 5.0 after 72 h of incubation. The best-suited substrate was nitrogen, and carbon sources were wheat bran, yeast, and lactose for enzyme production. In addition, metal ions Ca+2 and Mg+2 showed a net positive effect on the rate of the enzymatic reaction, and Al+3 and Co+2 inhibited the activity. Protein purification results demonstrated a 4-fold increase with 13% yield and specific activity of 40 U/mL/min. SDS-PAGE indicated the molecular weight of partially purified protein as ∼56 kDa. The stain removal efficiency of the purified enzyme showed considerable results. The finding revealed the use of low-cost and readily available substrates, making C. portucalensis (A-4) a promising candidate for detergent and pharmaceutical industries.
format Article
id doaj-art-6e1bcede132e4dbdbf167c233b7c94f9
institution DOAJ
issn 2307-4116
language English
publishDate 2024-01-01
publisher Elsevier
record_format Article
series Kuwait Journal of Science
spelling doaj-art-6e1bcede132e4dbdbf167c233b7c94f92025-08-20T03:07:26ZengElsevierKuwait Journal of Science2307-41162024-01-01511100156https://doi.org/10.1016/j.kjs.2023.11.005Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeuticsUme-e-Salma Liaqat0Shumaila Naz1Hafiz Iftikhar Hussain2Dept. of Bioscience, University of Wah, Wah Cantt, 47010, PakistanDept. of Bioscience, University of Wah, Wah Cantt, 47010, PakistanDept. of Pathology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur, 63100, PakistanStarch hydrolyzing amylases are among the vital enzymes used in the food, textile, detergent, and pharmaceutical industries. The aim was to isolate amylase bacteria from agricultural soil, its extraction and subsequent purification. Within the scope of this work, using a 1% starch agar medium, potent thermostable bacterial amylase (A-4) was isolated from agricultural soil samples. Molecular analysis revealed the amylase-producing bacterial strain (A-4) to be Citrobacter portucalensis. We used agro-industrial-based wastes (wheat bran, potato peel, and sugarcane bagasse) in amylase production. Culture condition optimization showed that high amylase activity of 4.96 U/mL was obtained at temperature 55 °C and pH 5.0 after 72 h of incubation. The best-suited substrate was nitrogen, and carbon sources were wheat bran, yeast, and lactose for enzyme production. In addition, metal ions Ca+2 and Mg+2 showed a net positive effect on the rate of the enzymatic reaction, and Al+3 and Co+2 inhibited the activity. Protein purification results demonstrated a 4-fold increase with 13% yield and specific activity of 40 U/mL/min. SDS-PAGE indicated the molecular weight of partially purified protein as ∼56 kDa. The stain removal efficiency of the purified enzyme showed considerable results. The finding revealed the use of low-cost and readily available substrates, making C. portucalensis (A-4) a promising candidate for detergent and pharmaceutical industries.https://www.sciencedirect.com/science/article/pii/S2307410823001979agro-industrial wasteamylasedetergentshydrolytic enzymeprotein purification
spellingShingle Ume-e-Salma Liaqat
Shumaila Naz
Hafiz Iftikhar Hussain
Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics
Kuwait Journal of Science
agro-industrial waste
amylase
detergents
hydrolytic enzyme
protein purification
title Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics
title_full Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics
title_fullStr Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics
title_full_unstemmed Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics
title_short Optimization and production of highly stable amylase isolated from Citrobacter portucalensis using low-cost agro-industrial wastes as substrates: Prospective therapeutics
title_sort optimization and production of highly stable amylase isolated from citrobacter portucalensis using low cost agro industrial wastes as substrates prospective therapeutics
topic agro-industrial waste
amylase
detergents
hydrolytic enzyme
protein purification
url https://www.sciencedirect.com/science/article/pii/S2307410823001979
work_keys_str_mv AT umeesalmaliaqat optimizationandproductionofhighlystableamylaseisolatedfromcitrobacterportucalensisusinglowcostagroindustrialwastesassubstratesprospectivetherapeutics
AT shumailanaz optimizationandproductionofhighlystableamylaseisolatedfromcitrobacterportucalensisusinglowcostagroindustrialwastesassubstratesprospectivetherapeutics
AT hafiziftikharhussain optimizationandproductionofhighlystableamylaseisolatedfromcitrobacterportucalensisusinglowcostagroindustrialwastesassubstratesprospectivetherapeutics