Screening of Lactic Acid Bacteria and RSM-Based Optimization for Enhancing γ-Aminobutyric Acid (GABA) Accumulation in Orange Juice - Research paper

Sep 30, 2026

Inoculated fermentation improves juice flavor, nutrition, and functional properties by using lactic acid bacteria (LAB) as starter cultures. This study screened 52 LAB strains for orange-juice fermentation and applied response surface methodology (RSM) to optimize key fermentation variables to raise γ-aminobutyric acid (GABA) accumulation. Two strains, Lacticaseibacillus paracasei ZY and Lacticaseibacillus rhamnosus SN12, produced higher GABA and showed strong substrate adaptability.

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CategoryHealthcare

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1. Introduction

Abstract

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1. Introduction (Received, Revised, Accepted, Published)

Article Screening of Lactic Acid Bacteria and RSM-Based Optimization for Enhancing γ-Aminobutyric Acid (GABA) Accumulation in Orange Juice Shufeng Yin, Yiyao Wang, RuiXue Zhao, Ning Zhao, Hao Liu, Yining Tang, Ningbo Qin , Yiwei Dai * and Xinping Lin * State Key Laboratory of Marine Food Processing & Safety Control, National Engineering Research Center of Seafood, School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China; [yinshufeng1220@163.com](yinshufeng1220@163.com) (S.Y.); [wangyiyao0506@163.com](wangyiyao0506@163.com) (Y.W.); [zhaorx04@outlook.com](zhaorx04@outlook.com) (R.Z.); [ningzhao0221@163.com](ningzhao0221@163.com) (N.Z.); [18763666325@163.com](18763666325@163.com) (H.L.); [18904084306@163.com](18904084306@163.com) (Y.T.); [qinningbo6902765@163.com](qinningbo6902765@163.com) (N.Q.) * [Correspondence: ywdai6228@126.com](Correspondence: ywdai6228@126.com) (Y.D.); [yingchaer@163.com](yingchaer@163.com) (X.L.); Tel.: +86-15242684612 (Y.D.); +86-15898160352 (X.L.) Abstract Inoculated fermentation can enhance the flavor, nutrition, and functionality of juice. The lactic acid bacteria (LAB) are commonly used as starter cultures. This study screened LAB for orange juice fermentation and optimized fermentation factors using response surface methodology (RSM) to improve GABA content in orange juice. A total of 52 LAB strains were screened, and Lacticaseibacillus paracasei ZY (Lcb. paracasei ZY) and Lacticaseibacillus rhamnosus SN12 (Lcb. rhamnosus SN12) presented higher GABA yields and adaptability to substrates. The optimized fermentation factors for GABA enhancement in orange juice were as follows: initial pH of 5.5, fermentation temperature of 37 ◦ C, soluble solids content of 12.0 ◦ Bx, inoculum ratio of Lcb. paracasei ZY to Lcb. rhamnosus SN12 as 1:1, inoculum size of 6 Log CFU/mL, and fermentation time of 96 h. Under these optimized conditions, the GABA content reached 0.89 g/L, representing a 39.06% increase compared to uninoculated orange juice. This indicates that RSM-based optimization is conducive to increasing GABA content in orange juice and provides a scientific basis for the development of GABAenriched functional fermented juices. Keywords: γ-aminobutyric acid (GABA); orange juice; Lacticaseibacillus paracasei; Lacticaseibacillus rhamnosus; single-factor experiment; response surface methodology (RSM); functional beverage 1. Introduction Academic Editor: Svetoslav Todorov Received: 22 November 2025 Revised: 9 December 2025 Accepted: 23 December 2025 Published: 25 December 2025 Copyright: © 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license. γ-aminobutyric acid (GABA) is a non-protein amino acid that is widely present in plants and animals. As a major inhibitory neurotransmitter in the central nervous system, GABA has multiple physiological functions, including improving sleep [1], alleviating anxiety [2], lowering blood pressure [3], and regulating blood sugar [4] . Compared with chemical synthesis and plant extraction methods, microbial fermentation is safer and more sustainable for GABA production [5] . Lactic acid bacteria (LAB), as generally recognized as safe (GRAS) probiotics, can irreversibly decarboxylate glutamic acid or monosodium glutamate (MSG) to convert GABA through glutamic acid decarboxylase (GAD) . In recent years, they have received increasing attention in the food industry [6] . The reported species of GABA-producing LAB include Levilactobacillus brevis (L. brevis) [7], Lactiplantibacillus plantarum (Lpb. plantarum) [8], Lacticaseibacillus rhamnosus (Lcb. rhamnosus) [9], Lacticaseibacillus paracasei (Lcb. paracasei) [10], and Limosilactobacillus fermentum (Lim. fermentum) [11] . Oranges are rich in nutrients, containing carbohydrates, vitamin C, and polyphenols [12] . Their hig

FAQs

How was GABA in orange juice enhanced in this study?

The work used inoculated LAB fermentation and then optimized fermentation conditions with response surface methodology (RSM) to increase GABA accumulation in orange juice.

Which LAB strains showed the best performance?

Lacticaseibacillus paracasei ZY and Lacticaseibacillus rhamnosus SN12 produced higher GABA yields and demonstrated better adaptability to the substrate.

What was the improvement in GABA content under optimized conditions?

Under optimized parameters, the GABA content reached 0.89 g/L, which was a 39.06% increase compared with uninoculated orange juice.

Sep 30, 2026

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