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108. Heterologous expression, fermentation strategies and molecular modification of collagen for versatile applications.

Author

Xiang, Zhi-Xiang, Gong, Jin-Song, Li, Heng, Shi, Wei-Ting, Jiang, Min, Xu, Zheng-Hong, and Shi, Jin-Song

Subjects
GENE expression, PROLINE hydroxylase, DEVELOPMENTAL biology, SYNTHETIC biology, MARINE mammals, COLLAGEN
Abstract

Collagen is a kind of high macromolecular protein with unique tissue distribution and distinctive functions in the body. At present, most collagen products are extracted from the tissues and organs of mammals or marine fish. However, this method exhibits several disadvantages, including low efficiency and serious waste generation, which makes it difficult to meet the current market demand. With the rapid development of synthetic biology and the deepening of high-density fermentation technology, the collagen preparation by biosynthesis strategy emerges as the times require. Co-expression with the proline hydroxylase gene can solve the problem of non-hydroxylated collagen, but the yield may be affected. Therefore, improving the expression through molecular modification and dynamic regulation of synthesis is an entry point for future research. Due to the defects in certain properties of the natural collagen, modification of properties would be benefit for meeting the requirements of practical application. In this paper, in-depth investigations on recombinant expression, fermentation, and modification studies of collagen are conducted. Also, it summarizes the research progress of collagen in food, medicine, and beauty industry in recent years. Furthermore, the future development trend and application prospect of collagen are discussed, which would provide guidance for its preparation and application. [ABSTRACT FROM AUTHOR]

Published
2023
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115. Biocatalysis

Author

Gong, Jin-Song, primary, Shi, Jin-Song, additional, and Xu, Zheng-Hong, additional

Published
2016
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116. Additional file 1 of High-efficiency secretory expression and characterization of the recombinant type III human-like collagen in Pichia pastoris

Author

Xiang, Zhi-Xiang, Gong, Jin-Song, Shi, Jin-Hao, Liu, Chun-Fang, Li, Heng, Su, Chang, Jiang, Min, Xu, Zheng-Hong, and Shi, Jin-Song

Abstract

Additional file 1: Table S1. List of primers involved in this work. Table S2. List of primers involved in RT-PCR. Fig. S1. SDS-PAGE analysis of 2# recombinant strain. Lanes 1-3 were the blank control (GS115/pPIC9K), lanes 4-6 were the 2# recombinant strain (GS115/pPIC9K-col), and lane M was standard molecular weights. Fig. S2. Target gene copy number of double-driving recombinant strain. Fig. S3. N-terminal sequencing results of the target protein. Fig. S4. C-terminal sequencing results of the target protein. Fig. S5. Western blotting analysis of the native hlCOLIII (after Native-PAGE). Lane 1 was the hlCOLIII, and lane M was standard molecular weights.

Published
2022
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124. Characterization of Cinnamomum kanehiraeExtract-Stimulated Triterpenoids Synthesis in Submerged Fermentation of Antrodia camphoratavia Untargeted Metabolomics

Author

Luo, Zhi-Shan, Lu, Zhen-Ming, Hu, Miao-Miao, Li, Xin-Yang, Xu, Guo-Qiang, Gong, Jin-Song, Shi, Jin-Song, and Xu, Zheng-Hong

Abstract

The underlying mechanisms of Cinnamomum kanehirae-stimulated growth and metabolism of Antrodia camphorataremain unknown. Herein, we first observed that the methanol extract of C. kanehiraetrunk (MECK) (2 g/L) showed a potent stimulatory effect on A. camphoratatriterpenoids production (115.6 mg/L). Second, MECK treatment considerably increased the category and abundance of many secondary metabolites in the mycelia. We identified 93 terpenoids (8 newly formed and 49 upregulated) in the MECK-treated mycelia, wherein 21 terpenoids were the same as those in the fruiting bodies. Third, 42 out of the 93 terpenoids were annotated in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, mainly involving monoterpenoids and diterpenoids syntheses. Finally, 27 monoterpenes and 16 sesquiterpenes were detected in the MECK, and the two terpenoids with the highest abundance (linalool and α-pinene) were selected for verification and found to considerably increase the terpenoids production of A. camphorataand demonstrate the regulation of mRNA expression levels of nine key genes in the mevalonate pathway via RT-qPCR. This study is beneficial for elucidating the terpenoids synthesis mechanism in A. camphorata.

Published
2023
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126. Desulfovibrio fairfieldensis -Derived Outer Membrane Vesicles Damage Epithelial Barrier and Induce Inflammation and Pyroptosis in Macrophages.

Author

Nie, Yawen, Xie, Xiao-Qian, Zhou, Lingxi, Guan, Qijie, Ren, Yilin, Mao, Yong, Shi, Jin-Song, Xu, Zheng-Hong, and Geng, Yan

Subjects
EXTRACELLULAR vesicles, TIGHT junctions, PYROPTOSIS, TUMOR necrosis factors, PARTICLE size determination, EPITHELIAL cells
Abstract

Sulfate-reducing bacteria Desulfovibrio fairfieldensis is an opportunistic pathogen that widely exists in the human intestine and can cause severe infectious diseases. However, the mechanisms contributing to its pathogenesis remain of great interest. In this study, we aim to investigate the outer membrane vesicles (OMVs) secreted by D. fairfieldensis and their pathogenic effect. The OMVs separated by ultracentrifugation were spherical and displayed a characteristic bilayer lipid structure observed by transmission electron microscopy, with an average hydrodynamic diameter of 75 nm measurement using the particle size analyzer. We identified 1496 and 916 proteins from D. fairfieldensis and its OMVs using label-free non-target quantitative proteomics, respectively. The 560 co-expressed proteins could participate in bacterial life activities by function prediction. The translocation protein TolB, which participates in OMVs biogenesis and transporting toxins was highly expressed in OMVs. The OMVs inhibited the expression of tight junction proteins OCCLUDIN and ZO-1 in human colonic epithelial cells (Caco-2). The OMVs decreased the cell viability of monocyte macrophages (THP-1-Mφ) and activated various inflammatory factors secretion, including interferon-γ (IFN-γ), tumor necrosis factor (TNF-α), and many interleukins. Further, we found the OMVs induced the expression of cleaved-gasdermin D, caspase-1, and c-IL-1β and caused pyroptosis in THP-1-Mφ cells. Taken together, these data reveal that the D. fairfieldensis OMVs can damage the intestinal epithelial barrier and activate intrinsic inflammation. [ABSTRACT FROM AUTHOR]

Published
2023
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143. Restricted intake of sulfur-containing amino acids reversed the hepatic injury induced by excess Desulfovibriothrough gut–liver axis

Author

Zhou, Lingxi, Lu, Gexue, Nie, Yawen, Ren, Yilin, Shi, Jin-Song, Xue, Yuzheng, Xu, Zheng-Hong, and Geng, Yan

Abstract

ABSTRACTDiet is a key player in gut–liver axis. However, the effect of different dietary patterns on gut microbiota and liver functions remains unclear. Here, we used rodent standard chow and purified diet to mimic two common human dietary patterns: grain and plant-based diet and refined-food-based diet, respectively and explored their impacts on gut microbiota and liver. Gut microbiota experienced a great shift with notable increase in Desulfovibrio, gut bile acid (BA) levels elevated significantly, and liver inflammation was observed in mice fed with the purified diet. Liver inflammation and elevated gut BA levels also occurred in mice fed with the chow diet after receiving Desulfovibrio desulfuricansATCC 29,577 (DSV). Restriction of sulfur-containing amino acids (SAAs) prevented liver injury mainly through higher hepatic antioxidant and detoxifying ability and reversed the elevated BA levels due to excess Desulfovibrio. Ex vivofermentation of human fecal microbiota with primary BAs demonstrated that DSV enhanced production of secondary BAs. Higher concentration of both primary and secondary BAs were found in the gut of germ-free mice after receiving DSV. In conclusion, Restriction of SAAs in diet may become an effective dietary intervention to prevent liver injury associated with excess Desulfovibrioin the gut.

Published
2024
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146. Coenzyme Q0 induces apoptosis, senescence and trans-differentiation of activated hepatic stellate cells

Author

Xu, Xin-Yi, Geng, Yan, Xu, Hong-Yu, Shi, Jin-Song, and Xu, Zheng-Hong

Abstract

Figure S1: Pathway functional enrichment of Coenzyme Q0 (7µM) vs control group. Bubble diagram showed KEGG pathway enrichment analysis. X-axis is enrichment ratio [Term Candidate Gene Number/Term Gene Number ], Y-axis is KEGG Pathway, the size of bubbles represents the number of genes annotated to KEGG Pathway, the color represents enrichment Q value, and the darker the color represents Q value, the smaller the value. Figure S2: Enrichment of DEGs in GO compared with the 7µM coenzyme Q0 treatment group. Gene ontology is divided into three major functional categories: molecular function (molecular function), cellular component (cellular component) and biological process (biological process). Perform functional classification based on the results of differential genetic testing. Under each major category, there are sub-categories of respective levels. Plot the GO annotation classification results of differential genes. The X axis represents the number of genes annotated into the GO cylinder, and the Y axis represents GO functional classification. Table S1: The expression levels of differentially expressed genes identified in three comparison groups ( CTL, 7μM CoQ0 treatment, 14μM CoQ0 treatment). Figure S1: Pathway functional enrichment of Coenzyme Q0 (7µM) vs control group. Bubble diagram showed KEGG pathway enrichment analysis. X-axis is enrichment ratio [Term Candidate Gene Number/Term Gene Number ], Y-axis is KEGG Pathway, the size of bubbles represents the number of genes annotated to KEGG Pathway, the color represents enrichment Q value, and the darker the color represents Q value, the smaller the value. Figure S2: Enrichment of DEGs in GO compared with the 7µM coenzyme Q0 treatment group. Gene ontology is divided into three major functional categories: molecular function (molecular function), cellular component (cellular component) and biological process (biological process). Perform functional classification based on the results of differential genetic testing. Under each major category, there are sub-categories of respective levels. Plot the GO annotation classification results of differential genes. The X axis represents the number of genes annotated into the GO cylinder, and the Y axis represents GO functional classification. Table S1: The expression levels of differentially expressed genes identified in three comparison groups ( CTL, 7μM CoQ0 treatment, 14μM CoQ0 treatment).

Published
2020
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