A New Approach to Produce Resveratrol by Enzymatic Bioconversion

Che, Jing; Shi, Junling; Gao, Zhenhong; Zhang, Yan

doi:10.4014/jmb.1512.12084

Cited by 12 publications

(12 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, keeping cells under starvation conditions might be helpful for the increase of resveratrol biosynthesis by Alternaria sp. MG1, which was consistent with experimental data [46].…”

Section: Resultssupporting

confidence: 92%

Genomic sequencing, genome-scale metabolic network reconstruction, and in silico flux analysis of the grape endophytic fungus Alternaria sp. MG1

Che

et al. 2019

Microb Cell Fact

Self Cite

View full text Add to dashboard Cite

BackgroundAlternaria sp. MG1, an endophytic fungus isolated from grape, is a native producer of resveratrol, which has important application potential. However, the metabolic characteristics and physiological behavior of MG1 still remains mostly unraveled. In addition, the resveratrol production of the strain is low. Thus, the whole-genome sequencing is highly required for elucidating the resveratrol biosynthesis pathway. Furthermore, the metabolic network model of MG1 was constructed to provide a computational guided approach for improving the yield of resveratrol.ResultsFirstly, a draft genomic sequence of MG1 was generated with a size of 34.7 Mbp and a GC content of 50.96%. Genome annotation indicated that MG1 possessed complete biosynthesis pathways for stilbenoids, flavonoids, and lignins. Eight secondary metabolites involved in these pathways were detected by GC–MS analysis, confirming the metabolic diversity of MG1. Furthermore, the first genome-scale metabolic network of Alternaria sp. MG1 (named iYL1539) was reconstructed, accounting for 1539 genes, 2231 metabolites, and 2255 reactions. The model was validated qualitatively and quantitatively by comparing the in silico simulation with experimental data, and the results showed a high consistency. In iYL1539, 56 genes were identified as growth essential in rich medium. According to constraint-based analysis, the importance of cofactors for the resveratrol biosynthesis was successfully demonstrated. Ethanol addition was predicted in silico to be an effective method to improve resveratrol production by strengthening acetyl-CoA synthesis and pentose phosphate pathway, and was verified experimentally with a 26.31% increase of resveratrol. Finally, 6 genes were identified as potential targets for resveratrol over-production by the recently developed methodology. The target-genes were validated using salicylic acid as elicitor, leading to an increase of resveratrol yield by 33.32% and the expression of gene 4CL and CHS by 1.8- and 1.6-fold, respectively.ConclusionsThis study details the diverse capability and key genes of Alternaria sp. MG1 to produce multiple secondary metabolites. The first model of the species Alternaria was constructed, providing an overall understanding of the physiological behavior and metabolic characteristics of MG1. The model is a highly useful tool for enhancing productivity by rational design of the metabolic pathway for resveratrol and other secondary metabolites.Electronic supplementary materialThe online version of this article (10.1186/s12934-019-1063-7) contains supplementary material, which is available to authorized users.

show abstract

“…Therefore, keeping cells under starvation conditions might be helpful for the increase of resveratrol biosynthesis by Alternaria sp. MG1, which was consistent with experimental data [46].…”

Section: Resultssupporting

confidence: 92%

Genomic sequencing, genome-scale metabolic network reconstruction, and in silico flux analysis of the grape endophytic fungus Alternaria sp. MG1

Che

et al. 2019

Microb Cell Fact

Self Cite

View full text Add to dashboard Cite

show abstract

“…As a newly characterized biological resource, Alternaria sp. MG1 is able to produce resveratrol without limitation from plant resources (Che et al 2016a; Shi et al 2012). Study of this fungus may allow insight into the necessary pathways allowing engineering for increased production.…”

Section: Discussionmentioning

confidence: 99%

Validation of reference genes for normalization of gene expression by qRT-PCR in a resveratrol-producing entophytic fungus (Alternaria sp. MG1)

Che

Shi

et al. 2016

AMB Expr

View full text Add to dashboard Cite

Alternaria sp. MG1, an endophytic fungus isolated from Vitis vinifera, can independently produce resveratrol, indicating that this species contains the key genes for resveratrol biosynthesis. Identification of these key genes is essential to understand the resveratrol biosynthesis pathway in this strain, which is currently unknown in microorganisms. qRT-PCR is an efficient and widely used method to identify the key genes related to unknown pathways at the level of gene expression. Verification of stable reference genes in this strain is essential for qRT-PCR data normalization, although results have been reported for other Alternaria sp. strains. In this study, nine candidate reference genes including TUBA, EF1, EF2, UBC, UFD, RPS5, RPS24, ACTB and 18S were evaluated for expression stability in a diverse set of six samples representing different growth periods. We compared cell culture conditions and an optimized condition for resveratrol production. The comparison of the results was performed using four statistical softwares. A combination of TUBA and EF1 was found to be suitable for normalization of Alternaria sp. MG1 in different developmental stages, and 18S was found to be the least stable. The reference genes verified in this study will facilitate further research to explore gene expression and molecular mechanisms as well as the improvement of secondary metabolite yields in Alternaria sp. MG1. To our knowledge, this is the first validation of reference genes in Alternaria with the capability to produce resveratrol. Additionally, these results provide useful guidelines for the selection of reference genes in other Alternaria species.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0283-z) contains supplementary material, which is available to authorized users.

show abstract

“…The recombinant enzyme used in repeated batch mode was able to transfer the sugar moiety from UDP‐glucose with up to 90% yield. Other examples on the use of enzymes for STBs production are provided by Che, Shi, Gao, and Zhang () who used immobilized enzymes from an Alternaria sp . grape endophytic fungus.…”

Section: Biotechnology and Genetic Engineering Approaches In Stbs Promentioning

confidence: 99%

Resveratrol and Related Stilbenoids, Nutraceutical/Dietary Complements with Health‐Promoting Actions: Industrial Production, Safety, and the Search for Mode of Action

Navarro

Pinilla

Ortiz

et al. 2018

Comp Rev Food Sci Food Safe

View full text Add to dashboard Cite

This paper reviews the potential of stilbenoids as nutraceuticals. Stilbenoid compounds in wine are considered key factors in health-promoting benefits. Resveratrol and resveratrol-related compounds are found in a large diversity of vegetal products. The stilbene composition varies from wine to wine and from one season to another. Therefore, the article also reviews how food science and technology and wine industry may help in providing wines and/or food supplements with efficacious concentrations of stilbenes. The review also presents results from clinical trials and those derived from genomic/transcriptomic studies. The most studied stilbenoid, resveratrol, is a very safe compound. On the other hand, the potential benefits of stilbene intake are multiple and are apparently due to downregulation more than upregulation of gene expression. The field may take advantage from identifying the mechanism of action(s) and from providing useful data to show evidence for specific health benefits in a given tissue or for combating a given disease.

show abstract

A New Approach to Produce Resveratrol by Enzymatic Bioconversion

Cited by 12 publications

References 21 publications

Genomic sequencing, genome-scale metabolic network reconstruction, and in silico flux analysis of the grape endophytic fungus Alternaria sp. MG1

Genomic sequencing, genome-scale metabolic network reconstruction, and in silico flux analysis of the grape endophytic fungus Alternaria sp. MG1

Validation of reference genes for normalization of gene expression by qRT-PCR in a resveratrol-producing entophytic fungus (Alternaria sp. MG1)

Resveratrol and Related Stilbenoids, Nutraceutical/Dietary Complements with Health‐Promoting Actions: Industrial Production, Safety, and the Search for Mode of Action

Contact Info

Product

Resources

About