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biosynthetic pathway of prescription cucurbitacin IIa and high-level production of key triterpenoid intermediates in engineered yeast and tobacco

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Plant Communications 2024年第6期5卷 154-172页

作者： Geng Chen Zhaokuan Guo Yanyu Shu Yan Zhao Lei Qiu Shaofeng Duan Yuan Lin Simei He Xiaobo Li Xiaolin Feng Guisheng Xiang Bo Nian Yina Wang Zhiyuan Li Chongkang Yang Yang Shi Yingchun Lu Guanze Liu Shengchao Yang Guanghui Zhang Bing Hao State Key Laboratory of Conservation and Utilization of Bio-resources in Yunnan The Key Laboratory of Medicinal Plant Biology of Yunnan ProvinceNational&Local Joint Engineering Research Center on Germplasms Innovation&Utilization of Chinese Medicinal Materials in Southwest ChinaYunnan Agricultural UniversityKunming 650201China Yunnan Characteristic Plant Extraction Laboratory KunmingYunnan 650106China

Cucurbitacin IIa is a triterpenoid isolated exclusively from Hemsleya plants and a non-steroidal anti-inflammatory drug that functions as the main ingredient of prescription Hemslecin capsules and tablets in *** biology provides new strategies for production of such valuable cucurbitacins at a large scale;however,the biosynthetic pathway of cucurbitacin IIa has been unknown,and the heterologous production of cucurbitacins in galactose medium has been expensive and low *** this study,we characterized the functions of genes encoding two squalene epoxidases(HcSE1-2),six oxidosqualene cyclases(HcOSC1-6),two CYP450s(HcCYP87D20 and HcCYP81Q59),and an acyltransferase(HcAT1)in cucurbitacin IIa biosynthesis by heterologous expression in Saccharomyces cerevisiae and Nicotiana *** achieved high-level production of the key cucurbitacin precursor 11-carbonyl-20b-hydroxy-Cuol from glucose in yeast via modular engineering of the mevalonate pathway and optimization of P450 expression *** resulting yields of 46.41 mg/l 11-carbonyl-20b-hydroxy-Cuol and 126.47 mg/l total cucurbitacin triterpenoids in shake flasks are the highest yields yet reported from engineered ***,production of 11-carbonyl-20b-hydroxy-Cuol by transient gene expression in tobacco resulted in yields of 1.28 mg/g dry weight in *** work reveals the key genes involved in biosynthesis of prescription cucurbitacin IIa and demonstrates that engineered yeast cultivated with glucose can produce high yields of key triterpenoid *** describe a low-cost and highly efficient platform for rapid screening of candidate genes and high-yield production of pharmacological triterpenoids.

关键词： cucurbitacin IIa oxidosqualene cyclase OSC 11-carbonyl-20b-hydroxy-Cuol Saccharomyces cerevisiae Nicotiana benthamiana Hemsleya chinensis

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Conserved Local Structural Motifs in Glycoside Hydrolase Families Facilitate the Discovery of Functional Enzymes

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Journal of Agricultural and Food Chemistry 2025年第19期73卷 2025 May 5页

作者： Liang, Yu-Peng Zhao, Ya-Lan Yin, Zhong-Wei Gong, Xiao-Wei Han, Xiu-Lin Wen, Meng-Liang National Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan Key Laboratory of Microbial Diversity in Southwest China Ministry of Education Yunnan Institute of Microbiology School of Life Sciences Yunnan University Yunnan Kunming650500 China R&D Center China Tobacco Yunnan Industrial Co. Ltd. Kunming650224 China

Glycoside hydrolases (GHs) are vital for natural glycoside biotransformation, especially in enhancing the pharmacological effects of natural products like ginsenosides. In this study, we collected 67 microbial-derived ginsenoside-hydrolyzing enzymes from nine GH families. Despite differences in global structures, the key residues surrounding substrate binding in GH1 and GH3 exhibit conserved structural motifs. Leveraging these motifs, five GH genes from Cellulosimicrobium were cloned, and three enzymes (Cbgl496, Cbgl516, Cbgl766) were characterized. Experimental results demonstrated that Cbgl516, Cbgl766, and Cbgl841 specifically catalyzed the hydrolysis of the β(1-6) glycosidic bond in the C-20 sugar chain of ginsenoside Rb1 to yield Rd. Cbgl496 selectively catalyzed the hydrolysis of β(1-2) glycosidic bonds in the oligosaccharide chains at the C-3 position of ginsenosides Rb1, Rb2, Rb3, and Rc, thereby directionally producing the minor ginsenosides Gy XVII, Compound O, Compound Mx1, and Compound Mc1. Structural analysis of 109,994 GH1/GH3 models from AlphaFold database revealed conserved residues across various organisms, emphasizing evolutionary conservation in the 3D structure of the catalytic core region despite sequence diversity. This study underscores the importance of conserved local structural motifs in GHs, offering insights for functional enzyme screening and understanding enzyme diversity and industrial applications. © 2025 American Chemical Society.

关键词： Hydrolysis

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A pair of transporters controls mitochondrial Zn^(2+) levels to maintain mitochondrial homeostasis

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Protein & Cell 2022年第3期13卷 180-202页

作者： Tengfei Ma Liyuan Zhao Jie Zhang Ruofeng Tang Xin Wang Nan Liu Qian Zhang Fengyang Wang Meijiao Li Qian Shan Yang Yang Qiuyuan Yin Limei Yang Qiwen Gan Chonglin Yang State Key Laboratory of Conservation and Utilization of Bio-resources in Yunnan and Center for Life Sciences School of Life SciencesYunnan UniversityKunming 650091China State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China

Zn^(2+)is required for the activity of many mitochondrial proteins,which regulate mitochondrial dynamics,apoptosis and ***,it is not understood how the proper mitochondrial Zn^(2+)level is achieved to maintain mitochondrial *** Caenorhabditis elegans,we reveal here that a pair of mitochondrion-localized transporters controls the mitochondrial level of Zn^(2+).We demonstrate that SLC-30A9/ZnT9 is a mitochondrial Zn^(2+)*** of SLC-30A9 leads to mitochondrial Zn^(2+)accumulation,which damages mitochondria,impairs animal development and shortens the life *** further identify SLC-25A25/SCaMC-2 as an important regulator of mitochondrial Zn^(2+)*** of SLC-25A25 suppresses the abnormal mitochondrial Zn^(2+)accumulation and defective mitochondrial structure and functions caused by loss of ***,we reveal that the endoplasmic reticulum contains the Zn^(2+)pool from which mitochondrial Zn^(2+)is *** findings establish the molecular basis for controlling the correct mitochondrial levels for normal mitochondrial structure and functions.

关键词： mitochondria Zn^(2+)transporter C.elegans ER-mitochondrial contact development

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ScRNA-seq and ST-seq in liver research

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Cell Regeneration 2023年第1期12卷 375-385页

作者： Jia He Chengxiang Deng Leonard Krall Zhao Shan State Key Laboratory of Conservation and Utilization of Bio‑resources in Yunnan and Center for Life Sciences School of Life SciencesYunnan UniversityKunming 650091China

Spatial transcriptomics,which combine gene expression data with spatial information,has quickly expanded in recent *** application of this method in liver research,our knowledge about liver development,regeneration,and diseases have been greatly *** this field is moving forward,a variety of problems still need to be addressed,including sensitivity,limited capacity to obtain exact single-cell information,data processing methods,as well as *** like single-cell RNA sequencing(scRNA-seq)are usually used together with spatial transcriptome sequencing(ST-seq)to clarify cell-specific gene *** this review,we explore how advances of scRNA-seq and ST-seq,especially ST-seq,will pave the way to new opportunities to investigate fundamental questions in liver ***,we will discuss the strengths,limitations,and future perspectives of ST-seq in liver research.

关键词： Liver development Liver regeneration Liver disease Single-cell RNA sequencing Spatial transcriptome sequencing

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Re-evaluating data quality of dog mitochondrial,Y chromosomal,and autosomal SNPs genotyped by SNP array

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Zoological Research 2016年第6期37卷 356-360页

作者： Newton O.OTECKO Min-Sheng PENG He-Chuan YANG Ya-Ping ZHANG Guo-Dong WANG State Key Laboratory of Genetic Resources and Evolution Yunnan Key Laboratory of Molecular Biology of Domestic AnimalsGermplasm Bank of Wild SpeciesKunming Institute of ZoologyChinese Academy of Sciences Kunming College of Life Science University of Chinese Academy of Sciences State Key Laboratory for Conservation and Utilization of Bio-Resources Yunnan University

Quality deficiencies in single nucleotide polymorphism （SNP） analyses have important implications. We used missingness rates to investigate the quality of a recently published dataset containing 424 mitochonddal, 211 Y chromosomal, and 160 432 autosomal SNPs generated by a semicustom Illumina SNP array from 5 392 dogs and 14 grey wolves. Overall, the individual missingness rate for mitochondrial SNPs was -43.8%, with 980 （18.1%）individuals completely missing mitochondrial SNP genotyping （missingness rate=l）. In males, the genotype missingness rate was -28.8% for Y chromosomal SNPs, with 374 males recording rates above 0.96. These 374 males also exhibited completely failed mitochondrial SNPs genotyping, indicative of a batch effect. Individual missingness rates for autosomal markers were greater than zero, but less than 0.5. Neither mitochondrial nor Y chromosomal SNPs achieved complete genotyping （locus missingness rate=0）, whereas 5.9% of autosomal SNPs had a locus missingness rate=l. The high missingness rates and possible batch effect show that caution and rigorous measures are vital when genotyping and analyzing SNP array data for domestic animals. Further improvements of these arrays will be helpful to future studies.

关键词： SNP array Dog Mitochondrial Ychromosomal Autosomal

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Individual and combined toxic effects of cypermethrin and chlorpyrifos on earthworm

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Journal of Environmental Sciences 2011年第4期23卷 676-680页

作者： Shiping Zhou Changqun Duan Wong Hang Gi Michelle Fazhong Yang Xuehua Wang Key Laboratory for Forest Resources Conservation and Use in the Southwest Mountains of China of Ministry of Education Southwest Forestry University Kunming 650224 China Laboratory for Conservation and Utilization of Bio-Resource & School of Life Science Yunnan University Kunming 650091 China

Toxicities were assessed for a pyrethroid （cypermethrin） and an organophosphate insecticide （chlorpyrifos） individually and in combination. A series of tests were conducted on different responses （acute, chronic, behavioral） of earthworms of species Eisenia fetida andrei in the ecological risk assessment of these pesticides. The results showed that the toxicity of the mixture of cypermethrin and chlorpyrifos was significantly higher than either of these pesticides individually, especially on the earthworm’s chronic responses. At a concentration of 5 mg/kg, the mixture caused significant reductions on the growth and reproduction rates of earthworms, but did not cause any significant effect when the individual was tested. The increase in toxicity of the pesticide mixture means that the use of toxicity data obtained exclusively from single-pesticide experiments may underestimate the ecological risk of pesticides that actually present in the field.

关键词： earthworms pesticide combined effects cypermethrin chlorpyrifos

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Melanin precursors mediated adaption to temperature changes in fungus and animal via inhibition of lipid-mediated ferroptosis

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Science China(Life Sciences) 2023年第8期66卷 1800-1817页

作者： Xiaoyu Yang Yonghong Chen Longlong Zhang Jiangbo He Qunfu Wu Shuhong Li Donglou Wang Jianghui Gou Zhuang Wu Keqin Zhang Shenghong Li Xuemei Niu State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan and Key Laboratory for Southwest Microbial Diversity of the Ministry of EducationYunnan UniversityKunming 650032China State Key Laboratory of Phytochemistry and Plant Resources in West China Kunming Institute of BotanyChinese Academy of SciencesKunming 650201China Kunming Key Laboratory of Respiratory Disease Kunming UniversityKunming 650214China

The discovery of biological activities of natural products plays a vital part in drug *** mechanism by which organisms respond to temperature changes via biosynthesis of natural products remained largely cryptic.A thermophilic fungus under cold stress turned black and accumulated a polyketide metabolite 1 and lipid *** in 1 caused melanin loss and accumulated extra lipid mass,unexpectedly leading to seriously damaged mitochondria diagnostic for *** analysis revealed that lipid mass induced by cold stress intensively increased ferroptosis risk and 1 functioned as cell wall reinforcer against mass lipid accumulation and as reactive oxygen species scavenger against lipid *** also found that melanin in mice lowered lipid level but enhanced animal resistance to cold *** with melanin precursors significantly increased mouse cell survival rate under cold *** results unveiled a metabolite-lipid-ferroptosis-cold relationship,which provided mechanistic insights into the functions of most common metabolites and into diseases related to cold *** findings opened a perspective for developing anti-cold and anti-ferroptosis therapeutics and agents.

关键词： melanin precursor lipid ferroptosis temperature change anti-cold

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Molecular evolution of stress-response gene Leptin in high-altitude Chinese snub-nosed monkeys (Rhinopithecus genus)

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Chinese Science Bulletin 2010年第36期55卷 4132-4135页

作者： WANG XiaoPing1,2, JIN Wei1,2, YU Li1,2 & ZHANG YaPing1,2 1 laboratory for conservation and utilization of bio-resource, yunnan University, Kunming 650091, China 2 state key laboratory of Genetic resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China Laboratory for Conservation and Utilization of Bio-resource Yunnan University Kunming China State Key Laboratory of Genetic Resources and Evolution Kunming Institute of Zoology Chinese Academy of Sciences Kunming China

Chinese snub-nosed monkeys (genus Rhinopithecus, subfamily Colobinae), including R. bieti, R. brelichi and R. roxellana, are well-known as the non-human primates with the highest known altitudinal distribution. They represent an interesting model organism of adaptation to the extreme environmental stresses. However, no study at the molecular level has yet been reported for the high-altitude adaptation in Chinese snub-nosed monkeys. Leptin, as an adipocyte-derived hormone, is believed to play an important role in energy homeostasis in adaptation to high altitude environments. In the present study, we sequenced and compared leptin sequences of the Chinese snub-nosed monkeys (R. bieti and R. roxellana) with their lowland close relative R. avunculus and other Colobines. Unexpectedly, no amino acid changes were observed in the 7 Colobinae species examined, including the 2 Chinese snub-nosed monkeys, indicating no difference in the evolutionary pattern of the Leptin gene between high-altitude monkeys and their lowland counterparts. In contrast to a previous finding of adaptive evolution of Leptin gene in plateau pikas, our study suggests that this gene may not have an important role in high-altitude adaptation of Chinese snub-nosed monkeys. Other nuclear genes associated with energy metabolism, or mitochondrial genes, are most likely to be involved the molecular mechanism underlying adaptation of these monkeys to cold and hypoxia associated with the highland environment.

关键词： Leptin基因金丝猴属瘦素基因压力反应分子进化高空鼻极端环境

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Deciphering the puzzles of dog domestication

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Zoological Research 2020年第2期41卷 97-104页

作者： Zhe Zhang Saber Khederzadeh Yan Li State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan School of Life SciencesYunnan UniversityKunmingYunnan 650091China State Key Laboratory of Genetic Resources and Evolution Germplasm Bank of Wild SpeciesKunming Institute of ZoologyChinese Academy of SciencesKunmingYunnan 650223China Kunming College of Life Science University of Chinese Academy of SciencesKunmingYunnan 650204China

The domestic dog,as a highly successful domestication model,is well known as a favored human *** its domestication history should provide great insight into our understanding of the prehistoric development of human culture and ***,investigation on the mechanisms underpinning the morphological and behavioral traits associated with canid domestication syndrome is of significance not only for scientific study but also for human medical *** development of a multidisciplinary canine genome database,which includes enormous omics data,has substantially improved our understanding of the genetic makeup of ***,we reviewed recent advances associated with the original history and genetic basis underlying environmental adaptations and phenotypic diversities in domestic dogs,which should provide perspectives on improving the communicative relationship between dogs and humans.

关键词： Dog Domestication history Adaptation Phenotypic diversity,Domestic legacy

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Expression of BSE-associated proteins in the CNS and lymphoreticular tissues of cattle and buffalo

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Science Bulletin 2016年第17期61卷 1377-1383页

作者： Hui Zhao Si-Qi Wang Li-Li Qing Lin-Lin Liu Ya-Ping Zhang State Key Laboratory for Conservation and Utilization of Bioresource Yunnan University Kunming 650091 China School of Life Science Yunnan University Kunming 650091 China State Key Laboratory of Genetic Resources and Evolution Kunming Institute of Zoology Chinese Academy of Sciences Kunming 650223 China

Bovine spongiform encephalopathy(BSE),which has been documented in 190,000 cases of BSE-infected cattle,to our knowledge,has not been reported in *** protein(PrP) is critical to susceptibility and development of ***,a new PrP-like protein,Shadoo(Sho),has been shown to have overlapping expression patterns and shared functions with ***,we hypothesize that differences in expression at the transcriptional level and/or the post-transcriptional level of the two genes may be associated with these specific differences between cattle and *** compared the relative mRNA expression of the prion protein gene(PRNP) and the Shadoo gene(SPRN) in 3% RNA samples using real-time *** also analyzed PrP/Sho protein from 96 samples by Western *** results demonstrated significantly lower PrP expression in the cerebellum,obex,mesenteric lymph node,and bronchial lymph node tissues,but higher relative expression of Sho in the cerebrum and spleen in buffalos compared with *** these results support our primary assumption,Sho and PrP expressions did not correlate with corresponding mRNA expression,suggesting that the biological modulations of both PrP and Sho proteins are at post-translational ***,positive correlations between PRNP and SPRN were found in cattle and buffalo cerebrum as well as in buffalo ***,negative correlations between PrP and Sho were detected in buffalo cerebellum and obex *** findings suggest that additional post-transcriptional studies are warranted to elucidate mechanisms behind prion diseases.

关键词： BSE PRNP SPRN Cattle Buffalo

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