Specific bile salt hydrolase genes in Lactobacillus plantarum AR113 and relationship with bile salt resistance

Wang, Guangqiang; Yu, Huaning; Feng, Xin; Tang, Haibo; Xiong, Zhiqiang; Xia, Yongjun; Ai, Lianzhong; Song, Xin

doi:10.1016/j.lwt.2021.111208

Cited by 20 publications

(17 citation statements)

References 26 publications

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“…The growth curves of AR113 wild‐type strain and Δbsh strains cultured in MRS were similar to those of corresponding strains cultured in MRS containing unconjugated bile acid, but were significantly different from that of ten strains cultured in MRS with conjugated bile salt. Moreover, the knockout of bsh 1 inhibited the growth of microorganisms in MRS with 3.0 g L −1 glyco‐conjugated bile acid, which was consistent with the result obtained by Wang 32 . We also found that the knock‐out of bsh 4 instead promoted the growth of microorganisms in MRS with 3.0 g L −1 conjugated bile acid.…”

Section: Discussionsupporting

confidence: 91%

“…Moreover, the knockout of bsh1 inhibited the growth of microorganisms in MRS with 3.0 g L −1 glycoconjugated bile acid, which was consistent with the result obtained by Wang. 32 We also found that the knock-out of bsh4 instead promoted the growth of microorganisms in MRS with 3.0 g L −1 conjugated bile acid. AR113 Δbsh1,3,2,4 cultured in MRS with glyco-or tauro-conjugated bile salts grew slowest, which fully proved that the knockout of four bsh was not conducive to the resistance of AR113 Δbsh1,3,2,4 to conjugated bile salts.…”

Section: Discussionmentioning

confidence: 56%

“…Bacterial strains and culture conditions All bacteria strains, plasmids, and primers used in this study are listed in Table 1. Lactiplantibacillus plantarum AR113 wild-type strain was isolated from pickle and performed whole genome sequencing and gene annotation by Shanghai Meiji Biological Co., Ltd Plasmid pLH01, pLdbsh4 and some single, double, and triple bsh knockout mutants of AR113 were constructed in our previous study, 32 while double and quadruple bsh knockout mutants (AR113 Δbsh2,4 and AR113 Δbsh1,3,2,4) were constructed in this study. All the above strains were stored in the Shanghai Engineering Research Center of Food Microbiology.…”

Section: Methodsmentioning

confidence: 99%

“…Construction of Δbsh strains AR113Δbsh2,4 and AR113Δbsh1,3,2,4 mutants were constructed with reference to the RecE/T-assisted CRISPR-Cas9 method. 31,32 Taking the construction of AR113 Δbsh1,3,2,4 as an example, the general process of knocking out bsh4 from AR113 Δbsh1,3,2 was shown in Fig. 1(A).…”

Section: Methodsmentioning

confidence: 99%

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Genes encoding bile salt hydrolase differentially affect adhesion of Lactiplantibacillus plantarumAR113

Wang

Tang

et al. 2021

J Sci Food Agric

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BACKGROUND: Adhesion is considered important for Lactiplantibacillus to persist in the human gut and for it to exert probiotic effects. Lactiplantibacillus plantarum contains a considerable number and variety of genes encoding bile salt hydrolases (bsh), but their effects on microbial adhesion remain poorly understood. To clarify the effects of four bsh on adhesion, we tried to knock out bsh (Δbsh) of L. plantarum AR113 using the CRISPR-Cas9 method, and compared the growth, auto-aggregation (R AA ), coaggregation (R CA ), surface hydrophobicity (A HC ) of AR113 wild-type and Δbsh strains and their adhesion abilities to HT29 cells. RESULTS: We first obtained the AR113 Δbsh1,3,2,4 strain with four bsh knocked out. Their growth was significantly slower than the wild-type strain cultured in De Man, Rogosa, and Sharpe medium (MRS) with 3.0 g L −1 glyco-or tauro-conjugated bile acid. Bsh had no significant effect on the growth of ten strains cultured in MRS, but Δbsh1 inhibited their growth when cultured in MRS containing 3.0 g L −1 sodium glycocholate, whereas Δbsh4 instead promoted their growth in MRS with 3.0 g L −1 sodium glycocholate and sodium taurocholate. R CA and R AA were linearly positive for all strains except AR113 Δbsh2,4, and A HC and R AA were negatively correlated for most strains excluding AR113 Δbsh2, with R AA = 6.38-25.05%, R CA = 5.17-9.22%, and A CH = 3.22-47.71%. The adhesion ability of ten strains cultured in MRS was higher than that of strains cultured in MRS with 3.0 g L −1 bovine bile, and it was related to bsh2.CONCLUSION: Bsh differentially affected the adhesion of AR113 series strains. This adds to the available information about substrate-gene-performance, and provides new information to enable engineering to regulate the colonization of Lactiplantibacillus.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 56%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Genes encoding bile salt hydrolase differentially affect adhesion of Lactiplantibacillus plantarumAR113

Wang

Tang

et al. 2021

J Sci Food Agric

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“…The screening of mutants was performed following the protocol as described previously with modifications ( Wang et al, 2021 ). The transformant colonies of L. plantarum XJ25 were inoculated into the MRS agar plates with the addition of erythromycin.…”

Section: Methodsmentioning

confidence: 99%

Functional Verification of the Citrate Transporter Gene in a Wine Lactic Acid Bacterium, Lactiplantibacillus plantarum

Yang

Zhao²,

Chen³

et al. 2022

Front. Bioeng. Biotechnol.

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Organic acid metabolism by lactic acid bacteria plays a significant role in improving wine quality. During this process, the uptake of extracellular organic acids by the transporters is the first rate-limiting step. However, up to now, there is very little published research on the functional verification of organic acid transporter genes in wine lactic acid bacteria. In this study, a predicted citrate transporter gene JKL54_04345 (citP) by protein homology analysis was knocked out using a CRISPR/Cas9-based gene-editing system, and then complemented using the modified pMG36e vectors in a major wine lactic acid bacterium, Lactiplantibacillus plantarum XJ25, to verify its function in citrate metabolism for the first time. The results showed that the gene knockout mutant XJ25-ΔcitP lost the ability to utilize citric acid, while the gene complement mutant XJ25-ΔcitP-pMG36ek11-citP fully recovered the ability of citric acid utilization. Meanwhile, citP knockout and complement barely affected the utilization of l-malic acid. These indicated that citP in L. plantarum functioned as a citrate transporter and was the only gene responsible for citrate transporter. In addition, two modified plasmid vectors used for gene supplement in L. plantarum showed distinct transcription efficiency. The transcription efficiency of citP in XJ25-ΔcitP-pMG36ek11-citP mutant was 4.01 times higher than that in XJ25-ΔcitP-pMG36ek-citP mutant, and the utilization rate of citric acid in the former was 3.95 times higher than that in the latter, indicating that pMG36ek11 can be used as a high-level expression vector in lactic acid bacteria.

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Enhanced resistance of Lactiplantibacillus plantarum by expression of albumin

Ding,

Wang,

Tang

et al. 2024

J Sci Food Agric

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BACKGROUNDHuman serum albumin (HSA) is the most abundant protein in plasma, playing crucial roles in regulating osmotic pressure and maintaining protein homeostasis. It is widely applied in the clinical treatment of various diseases. HSA can be purified from plasma or produced using recombinant DNA technology. Due to the improved efficiency and reduced costs, a growing body of research has focused on enhancing albumin production through bacterial strain overexpression. However, there have been few studies on the effect of albumin on the characteristics of the overexpressing‐strain itself, particularly stress resistance. In this study, we utilized Lactiplantibacillus plantarum (L. plantarum) AR113 as the expression host and successfully constructed the albumin overexpression strain AR113‐pLLY01 through gene editing technology. The successful expression of albumin was achieved and subsequently compared with the wild‐type strain AR113‐pIB184.RESULTSThe results demonstrated that the survival rate of AR113‐pLLY01 was also significantly better than that of AR113‐pIB184 after lyophilization. In addition, AR113‐pLLY01 exhibited a significantly better protective effect than AR113‐pIB184 at pH 3, indicating that albumin possesses a certain tolerance to acidic stress. At bile salt concentrations higher than 0.03%, both strains showed limited growth, but at a concentration of 0.02%, AR113‐pLLY01 had a significant protective effect.CONCLUSIONThis study suggest that albumin can improve strain tolerance, which has significant implications for future applications. © 2024 Society of Chemical Industry.

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Specific bile salt hydrolase genes in Lactobacillus plantarum AR113 and relationship with bile salt resistance

Cited by 20 publications

References 26 publications

Genes encoding bile salt hydrolase differentially affect adhesion of Lactiplantibacillus plantarumAR113

Genes encoding bile salt hydrolase differentially affect adhesion of Lactiplantibacillus plantarumAR113

Functional Verification of the Citrate Transporter Gene in a Wine Lactic Acid Bacterium, Lactiplantibacillus plantarum

Enhanced resistance of Lactiplantibacillus plantarum by expression of albumin

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