SCIENTIA HORTICULTURAE
·
2022
·
IF 4.342
Sun Yingjie,Zhang Xinyue,Jiang Yaping,Wang Jihan,Li Bingru,Zhang Xinhua,Li Xiaoan,Li Fujun
Ethylene response factors 2 (ERF2) belongs to the AP2/ERF transcription factor family and plays key roles in ethylene signal transduction . The roles of MdERF2 in apple ( Malus × domestica ) cuticular wax synthesis regulated by ethylene were studied in this work. Result shows that MdERF2 overexpression (ERF2-OE) up-regulated the expression levels of wax-related synthetic genes long-chain acyl-CoA synthetase 2 ( MdLACS2), Wax synthase1 ( MdWSD1), Eceriferum 4 ( MdCER4) and Eceriferum 6 ( MdCER6) , while has no effect on the transcription of Eceriferum 1 ( MdCER1) , and shows different regulation effects on the expression levels of mid-chain alkane hydroxylase 1 ( MdMAH1) between calli and fruit. Furthermore, different MdERF2 expression levels changed the apparent structure of the apple cuticular wax and positively regulated the total wax accumulation. ERF2-OE also increased alkane and alcohol production but decreased acid and ester contents. Thus, ethylene regulated the synthesis of apple cuticular wax at the transcriptional level through MdERF2, thereby altering the wax structure and composition.
研究领域:
植物学
·
分子生物学
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2025
·Vol. 26
·期 3
·1092
·
IF 4.9
Siqi Liu,Aolin Ma,Jie Li,Zhixuan Du,Longfei Zhu,Guanping Feng
Coumarins are natural products commonly found in plants and are typical allelopathic substances that strongly affect the growth of plants after being exudated from the root and help plants absorb Fe in cases of iron deficiency. Although coumarins have been found to have multiple effects, this understanding is still relatively limited. Here, we show that coumarin significantly promotes the elongation of the hypocotyl by enhancing cell elongation. Further research has found that coumarin increases the content of BR in plants by enhancing the expression of brassinosteroid (BR) synthesis genes. The effect of coumarin on promoting hypocotyl elongation is completely blocked by the mutation of the BR synthesis geneDEETIOLATED 2(DET2) or the co-addition of the BR synthesis inhibitor brassinazole (BRZ). Genetic analysis using Arabidopsis mutants showed that coumarin promoting hypocotyl elongation depends on the signaling pathway of the BRs. Overall, coumarin promotes elongation of the hypocotyl by increasing the synthesis of BRs in plants. These results provide us with new insights into the role of coumarins and offer strong theoretical support for the mechanisms of interactions between plants.
研究领域:
植物学
·
遗传学
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产品关联
DOI
PubMed
高影响因子
Biotechnology Journal
·
2015
·Vol. 11
·期 2
·228-237
·
IF 3.49
Tian Ma,Yuanjie Zhou,Xiaowei Li,Fayin Zhu,Yongbo Cheng,Yi Liu,Zixin Deng,Tiangang Liu
As a highly valued keto-carotenoid, astaxanthin is widely used in nutritional supplements and pharmaceuticals. Therefore, the demand for biosynthetic astaxanthin and improved efficiency of astaxanthin biosynthesis has driven the investigation of metabolic engineering of native astaxanthin producers and heterologous hosts. However, microbial resources for astaxanthin are limited. In this study, we found that the α-Proteobacterium Sphingomonas sp. ATCC 55669 could produce astaxanthin naturally. We used whole-genome sequencing to identify the astaxanthin biosynthetic pathway using a combined PacBio-Illumina approach. The putative astaxanthin biosynthetic pathway in Sphingomonas sp. ATCC 55669 was predicted. For further confirmation, a high-efficiency targeted engineering carotenoid synthesis platform was constructed in E. coli for identifying the functional roles of candidate genes. All genes involved in astaxanthin biosynthesis showed discrete distributions on the chromosome. Moreover, the overexpression of exogenous E. coli idi in Sphingomonas sp. ATCC 55669 increased astaxanthin production by 5.4-fold. This study described a new astaxanthin producer and provided more biosynthesis components for bioengineering of astaxanthin in the future. Graphical Limited microbial resources for astaxanthin limit its widespread applications in nutritional supplements and pharmaceuticals. Sphingomonas sp. ATCC 55669 can produce astaxanthin naturally. Whole-genome sequencing with gene functional confirmation identifies the discrete astaxanthin biosynthetic genes, and overexpressed related genes in E. coli can increase astaxanthin production. The new astaxanthin producer and biosynthesis components provide more possibilities for bioengineering of astaxanthin in the future.
研究领域:
微生物学
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生物工程
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产品关联
DOI
PubMed
高影响因子
FUNGAL GENETICS AND BIOLOGY
·
2022
·
IF 3.883
Jiazhen Qin,Jianwen Teng,Zhongyu Li,Ning Xia,Baoyao Wei,Li Huang
Similar to Pu-erh tea, Liupao tea is a post-fermented tea that is produced through natural fermentation by microorganisms. Penicillium citrinum is involved in multiple production processes of Liupao tea that can produce citrinin, a secondary metabolite with renal toxicity; however, the effect of P. citrinum on the quality of Liupao tea has not been investigated yet. Citrinin production is regulated by approximately 16 biosynthesis genes. However, little is known about the genetic background of citrinin in the complex Liupao tea system. In the present study, we cultured P. citrinum on potato dextrose agar and Liupao tea powder media and analyzed the changes of its nutritional components in Liupao tea. We selected six citrinin biosynthesis genes identified in Monascus exhibiting homology and high sequence similarity to those in P. citrinum and further analyzed the expression of citrinin biosynthesis genes in Liupao tea and the changes in citrinin yield. The results showed that the changes in nutritional components of Liupao tea were closely related to the growth and metabolism of P. citrinum and the quality of the tea. Decreases in the contents of soluble sugars (from 10.29% to 9.58%), soluble pectins (from 3.71% to 3.13%), free amino acids (from 3.84% to 3.14%), and tea polyphenols (from 22.84% to 18.78%) were noted. The Spearman’s correlation analysis indicated that P. citrinum growth can improve the tea quality to some extent. Quantitative real-time PCR demonstrated that ctnA gene was a positive regulator of citrinin production regardless of the culture medium used. ctnA and orf5 expressions greatly influenced the metabolism of citrinin by P. citrinum in Liupao tea. In conclusion, the citrinin biosynthesis genes, ctnA and orf5 , may be the promising targets for developing strategies to control P. citrinum infection and citrinin biosynthesis in Liupao tea.
研究领域:
食品科学
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微生物学
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分子生物学
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制茶技术
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2015
·Vol. 99
·期 14
·5825-5832
·
IF 3.337
Xu Dong-Bo,Ma Min,Deng Zi-Xin,Hong Kui
The type II polyketide synthase (PKS) natural product enterocin ( 1 ) was isolated from a mangrove-derived novel species Streptomyces qinglanensis 172205 guided by genome sequence, and its putative biosynthetic gene cluster was revealed. Its natural analogues 5-deoxyenterocin ( 2 ) and wailupemycin A–C ( 3–5 ) were also identified by tandem mass spectrometry. By feeding experiments with aryl acids, strain 172205 was proved to incorporate partial exogenous starter units into enterocin- and wailupemycin-based analogues, thus being a new and suitable microorganism for engineering unnatural enc -derived polyketide metabolites. In addition, biological assays indicated that enterocin showed obvious inhibitory activity against β-amyloid protein (Aβ 1-42 ) fibrillation and moderate cytotoxicity against HeLa and HepG2 for the first time.
研究领域:
微生物学
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基因组学
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天然产物化学
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神经生物学
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DOI
PubMed
高影响因子
Frontiers in Microbiology
·
2018
·
IF 4.019
Peng Qinying,Gao Guixi,Lü Jin,Long Qingshan,Chen Xuefei,Zhang Fei,Xu Min,Liu Kai,Wang Yemin,Deng Zixin,Li Zhiyong,Tao Meifeng
Streptomyces lividans is a suitable host for the heterologous expression of biosynthetic gene clusters (BGCs) from actinomycetes to discover “cryptic” secondary metabolites. To improve the heterologous expression of BGCs, herein we optimized S. lividans strain SBT5 via the stepwise integration of three global regulatory genes and two codon-optimized multi-drug efflux pump genes and deletion of a negative regulatory gene, yielding four engineered strains. All optimization steps were observed to promote the heterologous production of polyketides, non-ribosomal peptides, and hybrid antibiotics. The production increments of these optimization steps were additional, so that the antibiotic yields were several times or even dozens of times higher than the parent strain SBT5 when the final optimized strain, S. lividans LJ1018, was used as the heterologous expression host. The heterologous production of these antibiotics in S. lividans LJ1018 and GX28 was also much higher than in the strains from which the BGCs were isolated. S. lividans LJ1018 and GX28 markedly promoted the heterologous production of secondary metabolites, without requiring manipulation of gene expression components such as promoters on individual gene clusters. Therefore, these strains are well-suited as heterologous expression hosts for secondary metabolic BGCs. In addition, we successfully conducted high-throughput library expression and functional screening (LEXAS) of one bacterial artificial chromosome library and two cosmid libraries of three Streptomyces genomes using S. lividans GX28 as the library-expression host. The LEXAS experiments identified clones carrying intact BGCs sufficient for the heterologous production of piericidin A1, murayaquinone, actinomycin D, and dehydrorabelomycin. Notably, due to lower antibiotic production, the piericidin A1 BGC had been overlooked in a previous LEXAS screening using S. lividans SBT5 as the expression host. These results demonstrate the feasibility and superiority of S. lividans GX28 as a host for high-throughput screening of genomic libraries to mine cryptic BGCs and bioactive compounds.
研究领域:
微生物学
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合成生物学
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药物开发
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产品关联
DOI
PubMed
高影响因子
STAR Protocols
·
2023
·
IF 0
Xiaoqian Zeng,Shuliu Wang,Mindong Liang,Weishan Wang,Yue Jiang,Fei Xu,Leshi Liu,Hao Yan,Yaojun Tong,Lixin Zhang,Gao-Yi Tan
Summary Large biosynthetic gene cluster (BGC) cloning is important for discovering natural product-based drugs and remains challenging in high GC content microorganisms (e.g., Actinobacteria). Here, we present an in vitro CRISPR-Cas12a-mediated protocol for direct cloning of large DNA fragments. We describe steps for crRNA design and preparation, genomic DNA isolation, and CRISPR-Cas12a cleavage and capture plasmid construction and linearization. We then detail target BGC and plasmid DNA ligation and transformation and screening for positive clones. For complete details on the use and execution of this protocol, please refer to Liang et al. 1
研究领域:
分子生物学
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生物技术
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基因工程
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DOI
PubMed
PLoS Pathogens
·
2021
·Vol. 17
·期 6
·e1009656
·
IF 6.823
Lijuan Mei,Xuewen Wang,Ying Yin,Guirong Tang,Chengshu Wang
The exopolysaccharide galactosaminogalactan (GAG) has been well characterized in Aspergilli, especially the human pathogen Aspergillus fumigatus . It has been found that a five-gene cluster is responsible for GAG biosynthesis in Aspergilli to mediate fungal adherence, biofilm formation, immunosuppression or induction of host immune defences. Herein, we report the presence of the conserved GAG biosynthetic gene cluster in the insect pathogenic fungus Metarhizium robertsii to mediate either similar or unique biological functions. Deletion of the gene cluster disabled fungal ability to produce GAG on germ tubes, mycelia and appressoria. Relative to the wild type strain, null mutant was impaired in topical infection but not injection of insect hosts. We found that GAG production by Metarhizium is partially acetylated and could mediate fungal adherence to hydrophobic insect cuticles, biofilm formation, and penetration of insect cuticles. In particular, it was first confirmed that this exopolymer is responsible for the formation of appressorium mucilage, the essential extracellular matrix formed along with the infection structure differentiation to mediate cell attachment and expression of cuticle degrading enzymes. In contrast to its production during A . fumigatus invasive growth, GAG is not produced on the Metarhizium cells harvested from insect hemocoels; however, the polymer can glue germ tubes into aggregates to form mycelium pellets in liquid culture. The results of this study unravel the biosynthesis and unique function of GAG in a fungal system apart from the aspergilli species.
Fungal cell wall associated exopolymer GAG was first identified from Aspergillus niger decades ago and has been well characterized in terms of its biosynthetic mechanism, pathological and biological functions in Aspergillus fungi, especially the human pathogen A . fumigatus . With the accomplishment of diverse fungal genomes, the conserved GAG biosynthetic gene cluster is found to patchily distribute in fungal species beyond Aspergilli. However, until this study, the contribution of the clustered genes to GAG production has not been investigated in any non- Aspergillus fungi. We performed gene deletions and functional characterizations of GAG in an insect pathogenic fungus Metarhizium robertsii , and found that the partially acetylated GAG can be produced by the fungus to contribute to fungal virulence during topical infection of insect hosts. This exopolymer can be produced on germ tubes, mycelia and appressoria to mediate cell adherence, cuticle penetration, biofilm formation, mycelium aggregation in liquid culture and in particular, appressorium mucilaginous matrix formation. In addition, it was found that loss of GAG production could alter appressorium protein expressions including the enzymes involved in degrading insect cuticles. Our data advance the understanding of GAG biosynthesis and functions in an animal pathogenic fungus that is highly divergent from Aspergilli.
研究领域:
微生物学
·
分子生物学
·
病理学
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DOI
PubMed
高影响因子
CHEMBIOCHEM
·
2013
·Vol. 14
·期 12
·1468-1475
·
IF 3.74
Yuhai Jiang,Haoxin Wang,Chunhua Lu,Yanjiao Ding,Yaoyao Li,Yuemao Shen
Graphical A choirmaster is found: An orphan gene cluster containing a chorismatase gene was indentified in Streptomyces sp. LZ35. Characterization showed it to be responsible for the biosynthesis of cuevaene A, a 3-HBA-derived polyketide that shows moderate antibacterial activity against Gram-positive bacteria. Genome sequence analysis of Streptomyces sp. LZ35 has revealed a large number of secondary metabolite pathways, including one encoded in an orphan type I polyketide synthase gene cluster that contains a putative chorismatase/3-hydroxybenzoate synthase gene. Mutagenesis and comparative metabolic profiling led to the identification of cuevaene A as the metabolic product of the gene cluster, thus making it the first 3-HBA containing polyketide biosynthetic gene cluster described to date. Cuv10 was proven to be responsible for the conversion of chorismate into 3-HBA; Cuv18 is speculated to be responsible for the 6-hydroxylation of 3-HBA during polyketide chain elongation. Additionally, several pathway-specific regulatory factors that affect the production of cuevaene A were identified. Our results indicate that targeted inactivation of a gene followed by comparative metabolic profiling is a useful approach to identify and characterize cryptic biosynthetic gene clusters.
研究领域:
微生物学
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分子生物学
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天然产物化学
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DOI
高影响因子
Frontiers in Microbiology
·
2022
·
IF 6.064
Xu Jing,Wang Bo,Wang Ming-Qing,Gao Jian-Jie,Li Zhen-Jun,Tian Yong-Sheng,Peng Ri-He,Yao Quan-Hong
Organophosphate compounds are widely used in pesticides to control weeds, crop diseases, and insect pests. Unfortunately, these synthetic compounds are hazardous and toxic to all types of living organisms. In the present work, Escherichia coli was bioengineered to achieve methyl parathion (MP) degradation via the introduction of six synthetic genes, namely, opdS, pnpAS, pnpBS, pnpCS, pnpDS, and pnpES, to obtain a new transformant, BL-MP. MP and its subsequent decomposition intermediates were completely degraded by this transformant to enter the metabolites of multiple anabolic pathways. The MP-degraded strain created in this study may be a promising candidate for the bioremediation of MP and potential toxic intermediates.
研究领域:
生物技术
·
环境科学
·
微生物学
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DOI
PubMed
高影响因子
RSC Advances
·
2017
·Vol. 7
·期 20
·12185-12193
·
IF 3.108
Yuhua Zhang,Keming Jiang,Degang Qing,Bing Huang,Jiayi Jiang,Shumei Wang,Chunyan Yan
Camptotheca acuminata Decne (Nyssaceae) is a major natural source of the anticancer drug camptothecin (CPT) and its derivatives. The leaves and stems are two of the main regions that accumulate CPT in C. acuminata. In this study, we successfully isolated cambial meristematic cells (CMCs) and dedifferentiated cells (DDCs) from C. acuminata stems and leaves, identified their characteristic features using micrograph analysis, and assessed their hypersensitivity to γ-irradiation and zeocin. The growth and kinetics curves of CMCs and DDCs were also studied. Furthermore, the accumulation of CPT and 10-hydroxycamptothecin (HCPT)—a more potent and less toxic CPT derivative—was assessed by liquid chromatography, and the transcriptional levels of nine genes encoding key enzymes involved in CPT and HCPT biosynthesis was assessed using quantitative real-time polymerase chain reaction (PCR) in CMCs and DDCs. The results showed that CMCs induced were particularly hypersensitive to γ-irradiation and zeocin and presented with abundant and small vacuoles, whereas DDCs were not. The above morphological and physiological characteristics of CMCs were consistent with previous reports. B5 medium was determined as the best growth medium for CMCs and DDCs. The accumulation of CPT and HCPT was higher in CMCs than DDCs, and the expression of IPI, G10H, ASA1, TSB, TDC1, TDC2, and STR was significantly upregulated in CMCs compared with DDCs; HMGR2 and HMGR3 were downregulated in CMCs. We speculate that the high accumulation of CPT and HCPT in CMCs was due to the upregulation of these seven genes.
研究领域:
药理学
·
植物生物学
·
分子生物学
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产品关联
DOI
高影响因子
Nature Communications
·
2023
·Vol. 14
·期 1
·1-10
·
IF 16.6
Yue Qun,Meng Jie,Qiu Yue,Yin Miaomiao,Zhang Liwen,Zhou Weiping,An Zhiqiang,Liu Zihe,Yuan Qipeng,Sun Wentao,Li Chun,Zhao Huimin,Molnár István,Xu Yuquan,Shi Shuobo
Synthetic biology requires efficient systems that support the well-coordinated co-expression of multiple genes. Here, we discover a 9-bp nucleotide sequence that enables efficient polycistronic gene expression in yeasts and filamentous fungi. Coupling polycistronic expression to multiplexed, markerless, CRISPR/Cas9-based genome editing, we develop a strategy termed HACKing (Highly efficient and Accessible system by CracKing genes into the genome) for the assembly of multigene pathways. HACKing allows the expression level of each enzyme to be precalibrated by linking their translation to those of host proteins with predetermined abundances under the desired fermentation conditions. We validate HACKing by rapidly constructing highly efficient Saccharomyces cerevisiae cell factories that express 13 biosynthetic genes, and produce model endogenous (1,090.41 ± 80.92 mg L −1 squalene) or heterologous (1.04 ± 0.02 mg L −1 mogrol) terpenoid products. Thus, HACKing addresses the need of synthetic biology for predictability, simplicity, scalability, and speed upon fungal pathway engineering for valuable metabolites.
研究领域:
合成生物学
·
分子生物学
·
基因工程
·
代谢工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）