1.Research progress in key technologies for the development of Dendrobium officinale: from a rare and endangered species to a 10-billion-RMB-level industry.
Jing-Jing LIU ; Qiao-Xian YU ; Dong-Hong CHEN ; Ling-Shang WU ; Jin-Ping SI
China Journal of Chinese Materia Medica 2025;50(13):3670-3678
Dendrobium officinale(DO) is a traditional Chinese medicinal and edible plant, while it is critically endangered worldwide. This article, primarily based on the original research findings of the author's team and available articles, provides a comprehensive overview of the factors contributing to the endangerment of DO and the key technologies for the conservation, efficient cultivation, and value-added utilization of this plant. The scarcity of wild populations, low seed-setting rates, lack of endosperm in seeds, and the need for symbiosis with endophytic fungi for seed germination under natural conditions are identified as the primary causes for the rarity and endangerment of DO. Artificial seed production and tissue culture are highlighted as key technologies for alleviating the endangered status. The physiological and ecological mechanisms underlying the adaptation of DO to epiphytic growth are explored, and it is proposed that breaking the coupling of high temperature and high humidity is essential for preventing southern blight, a devastating affliction of DO. The roles of endophytic fungi in promoting the growth, improving the quality, and enhancing the stress resistance of DO are discussed. Furthermore, the integration of variety breeding, environment selection, and co-culture with endophytic fungi is emphasized as a crucial approach for efficient cultivation. The value-added applications of DO in pharmaceuticals, health foods, food products, and daily chemicals-particularly in the food and daily chemical industries-are presented as key drivers for a 10-billion-RMB-level industry. This systematic review offers valuable insights for the further development, utilization, and industrialization of DO resources, as well as for the broader application of conservation strategies for other rare and endangered plant species.
Dendrobium/microbiology*
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Endangered Species
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Seeds/microbiology*
;
Fungi/physiology*
2.Neuroprotective and antidiabetic lanostane-type triterpenoids from the fruiting bodies of Ganoderma theaecolum.
Jiaocen GUO ; Li YANG ; Luting DAI ; Qingyun MA ; Jiaoyang YAN ; Qingyi XIE ; Yougen WU ; Haofu DAI ; Youxing ZHAO
Chinese Journal of Natural Medicines (English Ed.) 2025;23(2):245-256
Eight previously undescribed lanostane triterpenoids, including five nortriterpenoids with 26 carbons, ganothenoids A-E (1-5), and three lanostanoids, ganothenoids F-H (6-8), along with 24 known ones (9-32), were isolated from the fruiting bodies of Ganodrma theaecolum. The structures of the novel compounds were elucidated using comprehensive spectroscopic methods, including electronic circular dichroism (ECD) and nuclear magnetic resonance (NMR) calculations. Compounds 1-32 were assessed for their neuroprotective effects against H2O2-induced damage in human neuroblastoma SH-SY5Y cells, as well as their inhibitory activities against protein tyrosine phosphatase 1B (PTP1B) and α-glucosidase. Compound 4 demonstrated the most potent neuroprotective activity against H2O2-induced oxidative stress by suppressing G0/G1 phase cell cycle arrest, reducing reactive oxygen species (ROS) levels, and inhibiting cell apoptosis through modulation of B-cell lymphoma 2 protein (Bcl-2) and Bcl-2 associated X-protein (Bax) protein expression. Compounds 26, 12, and 28 exhibited PTP1B inhibitory activities with IC50 values ranging from 13.92 to 56.94 μmol·L-1, while compound 12 alone displayed significant inhibitory effects on α-glucosidase with an IC50 value of 43.56 μmol·L-1. Additionally, enzyme kinetic analyses and molecular docking simulations were conducted for compounds 26 and 12 with PTP1B and α-glucosidase, respectively.
Humans
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Fruiting Bodies, Fungal/chemistry*
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Triterpenes/isolation & purification*
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Neuroprotective Agents/isolation & purification*
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Protein Tyrosine Phosphatase, Non-Receptor Type 1/metabolism*
;
Ganoderma/chemistry*
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Apoptosis/drug effects*
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Hypoglycemic Agents/isolation & purification*
;
Molecular Structure
;
alpha-Glucosidases/metabolism*
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Cell Line, Tumor
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Reactive Oxygen Species/metabolism*
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Oxidative Stress/drug effects*
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Hydrogen Peroxide/toxicity*
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Molecular Docking Simulation
3.(±)-Talapyrones A-F: six pairs of dimeric polyketide enantiomers with unusual 6/6/6 and 6/6/6/5 ring systems from Talaromycesadpressus.
Meijia ZHENG ; Xinyi ZHAO ; Chenxi ZHOU ; Hong LIAO ; Qin LI ; Yuling LU ; Bingbing DAI ; Weiguang SUN ; Ying YE ; Chunmei CHEN ; Yonghui ZHANG ; Hucheng ZHU
Chinese Journal of Natural Medicines (English Ed.) 2025;23(8):932-937
(±)-Talapyrones A-F (1-6), six pairs of dimeric polyketide enantiomers featuring unusual 6/6/6 and 6/6/6/5 ring systems, were isolated from the fungus Talaromyces adpressus. Their structures were determined by spectroscopic analysis and HR-ESI-MS data, and their absolute configurations were elucidated using a modified Mosher's method and electronic circular dichroism (ECD) calculations. (±)-Talapyrones A-F (1-6) possess a 6/6/6 tricyclic skeleton, presumably formed through a Michael addition reaction between one molecule of α-pyrone derivative and one molecule of C8 poly-β-keto chain. In addition, compounds 2/3 and 4/5 are two pairs of C-18 epimers, respectively. Putative biosynthetic pathways of 1-6 were discussed.
Polyketides/isolation & purification*
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Talaromyces/chemistry*
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Stereoisomerism
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Molecular Structure
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Circular Dichroism
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Pyrones/chemistry*
4.Establishment and application of a genetic operating system in Wickerhamomyces ciferrii for the synthesis of tetraacetyl phytosphingosine.
Liu LIU ; Zheng'an YIN ; Li PAN
Chinese Journal of Biotechnology 2025;41(1):397-415
Wickerhamomyces ciferrii (W.c), an unconventional heterothallic yeast species, is renowned for its high production of tetraacetyl phytosphingosine (TAPS). Due to its excellent performance in TAPS production, this study aimed to construct a genetic operating system of W.c to enhance the production of TAPS and to screen high-yielding strains by mutagenesis and genetic engineering, thus laying the foundation for further development of industrial production of sphingolipid metabolites. In this study, we selected two autonomous replication elements (CEN, 2μ) and mined 11 endogenous promoter elements to establish a genetic operating system in W. ciferrii. The overexpression of Syr2 and Lcb2 in the sphingolipid metabolism pathway significantly increased the production of TAPS. Meanwhile, we established a method for the identification of haploid mating types of W. ciferrii by combining RT-PCR and flow cytometry. Five strains of W. ciferrii with different mating types constructed from the standard diploid W. ciferrii ATCC 14091 were screened out. A-type haploid W.c 140 showcased the highest production of TAPS with a yield of 4.74 mg/g and a titer of 32.61 mg/L. Mutant strains W.c 140-A9 and W.c 140-A11 were induced by atmospheric pressure room temperature plasma mutagenesis. The recombinant strains W.c 140 OELcb2 and W.c 140 OESyr2 with overexpression were constructed with the genetic operating system established in this study. The TAPS yields of the mutant strains increased by 61.39% and 67.09%, respectively, compared with that of starting strain W.c 140. The recombinant strains cultured in the LCBNB medium achieved yields of 10.60 mg/g and 12.14 mg/g, respectively, representing 2.24 and 2.56 times of that in strain W.c 140. Moreover, the yields of the two recombinant strains were significantly higher than that of the diploid strain ATCC 14091. The genetic operating system and the haploid strain W.c 140 established in this study provide a basis for the subsequent establishment of genetic engineering tools for W. ciferrii.
Sphingosine/genetics*
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Saccharomycetales/metabolism*
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Genetic Engineering/methods*
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Promoter Regions, Genetic
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Metabolic Engineering/methods*
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Fungal Proteins/genetics*
5.Recent progress in ergothioneine of edible fungi.
Linlei YANG ; Zhenhui SHEN ; Xiangying LUO ; Rongping LI ; Rongchun LI
Chinese Journal of Biotechnology 2025;41(2):574-587
Ergothioneine is a natural antioxidant known for its potent anti-inflammatory and antioxidative properties. It has been applied in various sectors such as food, cosmetics, and pharmaceuticals. Edible fungi, both wild and cultivated, stand as the primary natural sources capable of synthesizing ergothioneine. This paper reviews the research progress in the content, physiological functions, extraction and detection methods, synthetic genes and pathways, mycelium fermentation, and engineering strain construction for ergothioneine production. The aim is to provide a comprehensive reference for advancing the research and industrial development related to ergothioneine in edible fungi.
Ergothioneine/isolation & purification*
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Fungi/genetics*
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Antioxidants/metabolism*
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Fermentation
6.Arbuscular mycorrhizal fungi improve physiological metabolism and ameliorate root damage of Coleus scutellarioides under cadmium stress.
Yanan HOU ; Fan JIANG ; Shuyang ZHOU ; Dingyin CHEN ; Yijie ZHU ; Yining MIAO ; Kai CENG ; Yifang WANG ; Min WU ; Peng LIU
Chinese Journal of Biotechnology 2025;41(2):680-692
Soil cadmium pollution can adversely affect the cultivation of the ornamental plant, Coleus scutellarioides. Upon cadmium contamination of the soil, the growth of C. scutellarioides is impeded, and it may even succumb to the toxic accumulation of cadmium. In this study, we investigated the effects of arbuscular mycorrhizal fungi (AMF) on the adaptation of C. scutellarioides to cadmium stress, by measuring the physiological metabolism and the degree of root damage of C. scutellarioides, with Aspergillus oryzae as the test fungi. The results indicated that cadmium stress increased the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and the content of malondialdehyde (MDA) and proline (Pro) within the cells of C. scutellarioides, but inhibited mycorrhizal infestation rate, root vigour and growth rate to a great degree. With the same cadmium concentration, the inoculation of AMF significantly improved the physiological indexes of C. scutellarioides. The maximum decrease of MDA content was 42.16%, and the content of secondary metabolites rosemarinic acid and anthocyanosides could be increased by up to 27.43% and 25.72%, respectively. Meanwhile, the increase of root vigour was as high as 35.35%, and the DNA damage of the root system was obviously repaired. In conclusion, the inoculation of AMF can promote the accumulation of secondary metabolites, alleviate root damage, and enhance the tolerance to cadmium stress in C. scutellarioides.
Cadmium/toxicity*
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Mycorrhizae/physiology*
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Plant Roots/drug effects*
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Soil Pollutants/toxicity*
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Stress, Physiological
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Superoxide Dismutase/metabolism*
7.Expression and biological characterization of anti-CD63 single-chain variable fragment antibody in Pichia pastoris.
Chinese Journal of Biotechnology 2025;41(4):1440-1454
To prepare antibodies that specifically recognize the conserved domain in the large extracellular loop of the CD63 protein, we expressed anti-CD63 single-chain variable fragment (scFv) antibody in Pichia pastoris in a secreted form. The purified expression product was found to bind specifically with CD63 protein and recognize CD63 on the surface of SK-MEL-28 cells. The variable region of the anti-CD63 monoclonal antibody in an anti-CD63-positive cell line was sequenced. The anti-CD63 scFv consisted of a variable heavy chain and a variable light chain linked by a flexible peptide was then designed. After codon optimization, the gene was synthesized and cloned into the expression plasmid pPICZα-A. The SacI-linearized plasmid was electroporated into P. pastoris X33, and 1% methanol were used to induce the expression of scFv. The fermentation supernatant was purified by Ni column. Anti-CD63 scFv was identified by SDS-PAGE and Western blotting, and its biological activities were analyzed by immunoblotting, immunofluorescence, cell-based ELISA, and flow cytometry. A P. pastoris strain capable of expressing and secreting anti-CD63 scFv was successfully obtained. The antibody had a molecular weight of approximately 30 kDa and specifically recognized CD63 protein. The expression of anti-CD63 scFv in P. pastoris paves the way for the production of anti-CD63 antibodies on a large-scale, which is undoubtedly an economical and effective way of antibody acquisition.
Single-Chain Antibodies/immunology*
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Humans
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Tetraspanin 30/immunology*
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Recombinant Proteins/immunology*
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Pichia/genetics*
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Saccharomycetales/metabolism*
8.Research progress in the fungal bioluminescence pathway.
Lei LÜ ; Ke CHENG ; Zhitao XU ; Shijie AN ; Dang XU ; Hao DU
Chinese Journal of Biotechnology 2025;41(7):2545-2558
The fungal bioluminescence pathway (FBP) catalyzes the oxidation of endogenous caffeic acid to produce green bioluminescence through an enzymatic cascade. Genetic engineering of FBP into plants creates autoluminescent specimens that circumvent the substrate limitations of conventional reporter systems. These transgenic plants serve dual functions as aesthetic displays and versatile biosensing platforms, enabling applications in real-time gene expression monitoring, continuous environmental surveillance, and non-invasive bioimaging, offering novel opportunities for horticultural production, environmental conservation, and bioengineering applications. This review synthesizes current advances in plant FBP engineering and explores how machine learning approaches can optimize autoluminescent phenotypes, thereby accelerating innovation in agricultural biotechnology, environmental sensing, and synthetic biology applications.
Fungi/genetics*
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Plants, Genetically Modified/metabolism*
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Genetic Engineering
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Biosensing Techniques
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Luminescent Measurements
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Caffeic Acids/metabolism*
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Luminescence
9.Progress in the heterologous synthesis of sesquiterpenes by Yarrowia lipolytica.
Liuhuan WANG ; Yuyao SHAO ; Xiaolin CUI ; Jiake YANG ; Yan QIAO ; Longxing XU ; Yuejia SUN ; Pengcai LIU ; Lijie YUAN ; Shuncheng LIU
Chinese Journal of Biotechnology 2025;41(9):3426-3447
Sesquiterpenes are natural terpenes containing 15 carbon atoms. They are widely used in the perfume, pharmaceutical, and biofuel industries due to their remarkable biological activities. The traditional production of sesquiterpenes relies on chemical synthesis or plant extraction, which has the disadvantages of low yields and waste of resources. The construction of microbial cell factories for the efficient synthesis of sesquiterpenes by means of synthetic biology provides a new option. In recent years, with the development of metabolic engineering and synthetic biology, the heterologous synthesis of a variety of sesquiterpenes has been successfully achieved by metabolic engineering of the oleaginous yeast, Yarrowia lipolytica. In this paper, we review the research progress in the heterologous synthesis of different sesquiterpenes by Y. lipolytica, discuss the synthetic biology strategies commonly used in this field, and make an outlook on the research directions and engineering approaches to further enhance the sesquiterpene yield in this host. This paper provides a reference for strategies such as synergistic optimization of synthetic biology and metabolic engineering, enhanced precursors, and opens up new directions for the application of synthetic biology in green chemistry and sustainable production.
Yarrowia/genetics*
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Sesquiterpenes/metabolism*
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Metabolic Engineering/methods*
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Synthetic Biology/methods*
10.Signature motif identification and enzymatic characterization of a protein tyrosine phosphatase in Metarhizium anisopliae.
Ze TAN ; Pei ZHU ; Zhenlun LI ; Shuiying YANG
Chinese Journal of Biotechnology 2025;41(9):3579-3588
Protein tyrosine phosphatases (PTPs, EC 3.1.3.48) are key regulators of cellular processes, with the catalytic activity attributed to the conserved motif (H/V)CX5R(S/T), where cysteine and arginine residues are critical. Previous studies revealed that alternative splicing of extracellular phosphatase mRNA precursors in Metarhizium anisopliae generated two distinct transcripts, with the longer sequence containing a novel HCPTPMLS motif resembling PTP signatures but lacking the arginine residue. To identify the novel signature motif and characterize its enzymatic properties, we heterologously expressed and purified both proteins in Pichia pastoris and comprehensively characterized their enzymatic properties. The protein containing the HCPTPMLS motif (designated as L-protein) exhibited the highest activity at pH 5.5 and a strong preference for pTyr substrates. Its phosphatase activity was inhibited by Ag+, Zn2+, Cu2+, molybdate, and tungstate, but enhanced by Ca2+ and EDTA. AcP101 (lacking HCPTPMLS) showed the maximal activity at pH 6.5 and a strong preference toward pNPP (P < 0.05), with the activity inhibited by NaF and tartrate, but enhanced by Mg2+ and Mn2+. Functional analysis confirmed that the L-protein retained the PTP activity despite the absence of arginine in its signature motif, while AcP101 functioned as an acid phosphatase. This study provides the first functional validation of an arginine-deficient PTP motif, expanding the definition of PTP signature motifs and offering new insights for phosphatase classification.
Metarhizium/genetics*
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Protein Tyrosine Phosphatases/chemistry*
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Amino Acid Motifs
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Recombinant Proteins/biosynthesis*
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Amino Acid Sequence
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Pichia/metabolism*
;
Fungal Proteins/chemistry*
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Substrate Specificity
;
Saccharomycetales

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