A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification

Cold-adapted pectin lyases are particularly useful in the extraction and clarification of freshly squeezed fruit juices at low temperatures, as they effectively reduce juice viscosity and improve light transmittance. With the increasing attention on low-temperature pectinase in industrial applicatio...

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Main Authors: Yujing Bai, Jin Wang, Yongliang Yan, Yuhua Zhan, Zhengfu Zhou, Min Lin
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/3/634
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author Yujing Bai
Jin Wang
Yongliang Yan
Yuhua Zhan
Zhengfu Zhou
Min Lin
author_facet Yujing Bai
Jin Wang
Yongliang Yan
Yuhua Zhan
Zhengfu Zhou
Min Lin
author_sort Yujing Bai
collection DOAJ
description Cold-adapted pectin lyases are particularly useful in the extraction and clarification of freshly squeezed fruit juices at low temperatures, as they effectively reduce juice viscosity and improve light transmittance. With the increasing attention on low-temperature pectinase in industrial applications, the exploration of low-temperature pectinase with novel characteristics has become one of the key focuses of research and development. In this study, a 1026 bp gene, <i>pel1Ba</i>, encoding a 42.7 kDa pectin lyase, was cloned from sediment samples collected from the South China Sea and heterologously expressed in <i>Escherichia coli.</i> The purified Pel1Ba exhibited an optimal temperature of 40 °C and an optimal pH of 10, with a total enzyme activity of 5100 U/mL. Notably, Pel1Ba is a cold-adapted enzyme that retains 80% of its relative activity across the temperature range of 0–40 °C. When 20 U/mL purified Pel1Ba was added to orange juice, the juice volume increased by 43.00% and its clarity improved by 37.80%. Meanwhile, site-directed mutagenesis analysis revealed that the residual enzyme activities of the mutants A230I, F253I, and L292I were increased by 22.5%, 34.4%, and 25.1%, respectively, compared to the wild type. This study concludes that the cold-active pectate lyase Pel1Ba exhibits potential for applications in the food industry.
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spelling doaj-art-d1a17a316b8b4f1b95ffd59d024d7d742025-08-20T02:42:27ZengMDPI AGMicroorganisms2076-26072025-03-0113363410.3390/microorganisms13030634A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice ClarificationYujing Bai0Jin Wang1Yongliang Yan2Yuhua Zhan3Zhengfu Zhou4Min Lin5School of Life and Health Sciences, Hainan University, Haikou 570228, ChinaNational Key Laboratory of Agricultural Microbiology, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaNational Key Laboratory of Agricultural Microbiology, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaNational Key Laboratory of Agricultural Microbiology, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaNational Key Laboratory of Agricultural Microbiology, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, ChinaSchool of Life and Health Sciences, Hainan University, Haikou 570228, ChinaCold-adapted pectin lyases are particularly useful in the extraction and clarification of freshly squeezed fruit juices at low temperatures, as they effectively reduce juice viscosity and improve light transmittance. With the increasing attention on low-temperature pectinase in industrial applications, the exploration of low-temperature pectinase with novel characteristics has become one of the key focuses of research and development. In this study, a 1026 bp gene, <i>pel1Ba</i>, encoding a 42.7 kDa pectin lyase, was cloned from sediment samples collected from the South China Sea and heterologously expressed in <i>Escherichia coli.</i> The purified Pel1Ba exhibited an optimal temperature of 40 °C and an optimal pH of 10, with a total enzyme activity of 5100 U/mL. Notably, Pel1Ba is a cold-adapted enzyme that retains 80% of its relative activity across the temperature range of 0–40 °C. When 20 U/mL purified Pel1Ba was added to orange juice, the juice volume increased by 43.00% and its clarity improved by 37.80%. Meanwhile, site-directed mutagenesis analysis revealed that the residual enzyme activities of the mutants A230I, F253I, and L292I were increased by 22.5%, 34.4%, and 25.1%, respectively, compared to the wild type. This study concludes that the cold-active pectate lyase Pel1Ba exhibits potential for applications in the food industry.https://www.mdpi.com/2076-2607/13/3/634Pel1Balow-temperature-activejuice clarificationmarine bacterium
spellingShingle Yujing Bai
Jin Wang
Yongliang Yan
Yuhua Zhan
Zhengfu Zhou
Min Lin
A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification
Microorganisms
Pel1Ba
low-temperature-active
juice clarification
marine bacterium
title A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification
title_full A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification
title_fullStr A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification
title_full_unstemmed A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification
title_short A Low-Temperature-Active Pectate Lyase from a Marine Bacterium for Orange Juice Clarification
title_sort low temperature active pectate lyase from a marine bacterium for orange juice clarification
topic Pel1Ba
low-temperature-active
juice clarification
marine bacterium
url https://www.mdpi.com/2076-2607/13/3/634
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