APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2025
·Vol. 109
·期 1
·1-11
·
IF 4.3
Wang Lihong,Tian Xueqin,Xue Pinghong,Deng Yunhong,Gao Rui,Hu Zhihong
Ergothioneine (EGT) is a rare amino acid with potent antioxidant and anti-inflammatory properties, with a wide range of applications in food, cosmetics, and medicine. In the present study, Aspergillus oryzae , a common edible fungus, was engineered as an optimal host for EGT production. Moreover, two endogenous genes involved in EGT biosynthesis were characterized. The homolog AoEgt1 was shown to be localized in the vacuoles, whereas the homolog AoEgt2 was found in the peroxisomes. Overexpression of EGT biosynthetic genes from different organisms enhanced EGT production, yielding 15.17 mg EGT/g of dry weight. Using glucose as the carbon source and supplementing methionine (Met) as a precursor further increased EGT production to 20.03 mg EGT/g of dry weight, constituting an eight-fold increase compared to the wild-type strain. This study discusses the successful construction of a high-yielding A. oryzae strain for EGT biosynthesis, providing a novel strategy for efficient EGT synthesis. Key points • Two newly described homologs, AoEgt1 and AoEgt2, were identified in A. oryzae. • AoEgt1 and AoEgt2 were found to contribute to EGT biosynthesis. • EGT production was significantly increased by overexpression of Egt1 and Egt2. • Glucose and Met supplementation in the medium increased EGT production.
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BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
·
2022
·Vol. 27
·期 3
·443-450
·
IF 3.386
Yang Rong-ling,Chen Xi,Song Yu-ye,Zhu Qian-lin,Bilal Muhammad,Wang Yu,Tong Zheng,Wu Ting-ting,Wang Zhao-yu,Luo Hong-zhen,Zhao Xiang-jie,He Ting-ting
Tyrosinase inhibitors are clinically effective for treating some dermatological disorders related to melanin hyperpigmentation. Accordingly, the discovery and development of tyrosinase inhibitors have great value in the pharmaceutical and cosmetic industry. Here, a novel tyrosinase inhibitor, 6′-O-cinnamoyl-helicid (helicid cinnamylate) was successfully synthesized by a simple and effective biocatalytic approach with Aspergillus oryzae cells. Investigation of the effects of several key variables on helicid cinnamylate synthesis found that the reaction conversion, reaction rate and regioselectivity reached 99%, 9.40 mM/h and > 99%, respectively, at the optimal conditions with anhydrous acetone as the solvent, whole-cell concentration of 40 mg/mL, and the molar ratio of vinyl cinnamate to helicid of 10 at 45°C. The whole-cells retained 68.87% of its initial activity after reusing for seven batches, indicating a potent application potential in non-aqueous biocatalytic systems. It was worth noting that helicid cinnamylate demonstrated a more potent tyrosinase inhibitory activity with an IC 50 value of 3.55 mM than helicid (IC 50 = 4.48 mM) and arbutin (IC 50 = 5.48 mM), which suggest that helicid cinnamylate could be developed as a more potential tyrosinase inhibitor. In conclusion, this study provides a novel whole-cell catalytic approach for the synthesis of helicid cinnamylate and insight into its application as a tyrosinase inhibitor.
研究领域:
生物化学
·
药学
·
化妆品科学
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JOURNAL OF FOOD SCIENCE
·
2016
·Vol. 81
·期 8
·C1883
·
IF 1.649
Ning Xu,Yaqi Liu,Yong Hu,Mengzhou Zhou,Chao Wang,Dongsheng Li
en The autolyzed mycelia of Aspergillus oryzae are rich in proteins, nucleic acids, sugar, and other biomacromolecules, and are one of the main contributors to the flavor profile of commercially important fermented goods, including soy sauce and miso. We induced autolysis of the mycelia of A. oryzae over 1 to 10 d, and found that the maximum dissolved amounts of total protein and nucleic acid ratio accounted for 28.63% and 88.93%, respectively. The organic acid content, such as citric acid, tartaric acid, succinic acid, lactic acid, and acetic acid, initially increased and then decreased as autolysis progressed, corresponding to changes in pH levels. The main characteristic flavor compounds in soy sauce, namely, ethanol, 2-phenylethanol, and 2-methoxy-4-vinylphenol, were all detected in the autolysate. Subsequently, we tested the effect of adding mycelia of A. oryzae during the fermentation process of soy sauce for 60 d, and found that addition of 1.2‰ A. oryzae mycelia provided the richest flavor. Overall, our findings suggest that compounds found in the autolysate of A. oryzae may promote the flavor compounds of soy sauce, such as alcohols, aldehydes, phenols, and esters. Practical Application pt Autolysate of Aspergillus oryzae mycelium contains rich nutrients and volatile flavor compounds, which is an important component of fermentation seasoning. This study provides a theoretical basis for developing the autolysate products of mycelia of A. oryzae and studying the flavor formation of soy sauce. The autolysate of A. oryzae may also be used to prepare healthy drinks and seasoning products.
研究领域:
食品科学
·
微生物学
·
风味化学
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CHIRALITY
·
2019
·Vol. 32
·期 2
·231-238
·
IF 1.927
Peng Wu,Mengjie Zhang,Yinjun Zhang,Zhao Wang,Jianyong Zheng
In this study, a novel lipase M5 derived from Aspergillus oryzae WZ007 was prone to exhibit high hydrolytic activity and stereoselectivity towards racemic substrate ( R , S )-ethyl 2-bromoisovalerate. ( R )-ethyl 2-bromoisovalerate was obtained by enzymatic resolution, which is the key chiral intermediate for highly efficient enantiomerically fluvalinate. The results showed that the enzymatic reaction was carried out in 120mM racemic substrate for 3 hours, the enantiomeric excess reached 98.6%, the conversion was 51.7%, and E value above 120. Therefore, the novel lipase M5 has the ability to efficiently produce ( R )-ethyl 2-bromoisovalerate, which greatly reduces the industrial production cost of the highly efficient counterpart of fluvalinate.
研究领域:
生物化学
·
工业微生物学
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PROTEIN EXPRESSION AND PURIFICATION
·
2018
·
IF 1.338
Jian-yong Zheng,Xing Lan,Xiao-jun Li,Li-juan Huang,Yin-jun Zhang,Zhao Wang
Pichia pastoris expression is a mature and efficient eukaryotic expression system . In this work, Aspergillus oryzae lipase (AOL, with the molecular mass of 28 kDa), which can perform highly stereoselective hydrolysis of (R, S)-methyl 2-(4-hydroxyphenoxy) propanoate, was expressed in P . pastoris X-33. The specific activity of AOL was 432 U/mg, which was obtained by fed-batch cultivation in a 5 L bioreactor using a methanol feeding strategy. After fermentation, the supernatant was concentrated by ultrafiltration with a 10 kDa cut-off membrane and purified with DEAE-Sepharose™ FF ion-exchange chromatography and phenyl Seflnose™ 6 FF hydrophobic interaction chromatography . The purified lipase activity reached 5509 U/mg. AOL showed high activity toward short-chain triacylglyceride (C 4 ), and the optimum temperature and pH were 40 °C and 8.0, respectively. The purified enzyme activity was inhibited by Zn 2+ and Cu 2+ . Moreover, the K m and V max values were 1 mM and 32.89 mmol/min, respectively.
研究领域:
生物技术
·
酶学
·
微生物学
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Foods
·
2026
·Vol. 15
·期 10
·1753
·
IF 6
Ting Zhao,Yibing He,Ying Han,Qinhao Liu,Xinqi Jian,Wei Zhao,Chiyue Zhang,Xianbing Xu,Yiying Nian,Zhenyu Wang,Ming Du,Peng Liu,Liming Sun
In our preliminary work, a clam sauce prepared by fermentation withAspergillus oryzae3.042 (AO) exhibited desirable flavor and quality; however, the process was prolonged (exceeding 30 d), and a high salt concentration (6–15%) was necessary to prevent spoilage. Consequently, shortening production cycle and reducing salt content without compromising product quality became a new objective. Enzymatic hydrolysis has long been recognized as an efficient approach in seasoning production, with enzyme efficacy being a key competitive factor. Accordingly, an AO-derived aminopeptidase–protease complex (AOAP) was optimized and prepared as a preparatory step. In this study, AOAP was applied to hydrolyze clam meat to evaluate its potential for producing a seasoning base. A two-step enzymatic hydrolysis process was employed. In the first step, the highest hydrolysis degree (29.1%) was achieved using alkaline protease (AP). The resulting hydrolysate was subsequently subjected to secondary hydrolysis with AOAP, achieving a degree of hydrolysis as high as 49.8%. Sensory evaluation revealed a significant reduction in bitterness and enhancement of umami in the final hydrolysate, a finding corroborated by electronic tongue analysis. Further characterization via LC-MS and amino acid (aa) analysis showed that a substantial number of bitter and umami peptides were released following AP treatment; however, the number of these peptides was markedly reduced after a subsequent AOAP hydrolysis, with concurrent substantial changes in the peptide profile. In the two-step hydrolysate, umami peptides mostly contain 3–10 aa, whereas bitter peptides typically contain only 3–5 aa. The content of free aa increased from 369.17 mg/100 g in the control to 3026.25 mg/100 g in the two-step hydrolysate, half of which were bitter, indicating the debittering efficiency of AOAP. Electronic nose analysis revealed similar flavor profile and characteristic presence of nitrogen oxides in all hydrolysates. GC-MS analysis further demonstrated that, after combined enzymatic hydrolysis, the short-chain aldehydes and ketones responsible for the fishy odor in the raw material almost completely disappeared, while long-chain aldehydes with pleasant aromas were generated. These findings suggest that the secondary hydrolysis step using AOAP can effectively improve the overall flavor profile of the clam hydrolysate, which may support its potential applicability in seasoning production, though further optimization and scale-up validation are needed.
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JOURNAL OF ENVIRONMENTAL MANAGEMENT
·
2022
·
IF 8.91
Kaidi Hu,Jianlong Li,Tianye Zhao,Qiao Zhou,Qin Li,Xinjie Hu,Guoquan Han,Shuhong Li,Likou Zou,Shuliang Liu
As a major intermediate metabolite of synthetic pyrethroids , the occurrence of 3-phenoxybenzoic acid hinders the decomposition of the parent pesticide and poses uncertain risks to environmental ecology and living organisms. Strain Aspergillus oryzae M-4 was previously reported to degrade 3-PBA and several substances were identified as downstream transformation products (TPs). But the mechanism underlying the cleavage of ether bond remains largely unclear. Here, we attempted to address such concern through identifying the peripheral TPs and analyzing transcriptomics, coupled with serial batch degradation experiments. Analysis results of chromatographic/mass spectrometry suggested that 3-PBA underwent twice hydroxylation , to yield mono- and dihydroxylated 3-PBA successively. In parallel, a mutual transformation between 3-PBA and 3-phenoxybenzyl alcohol (3-PBOH) also existed. The proposal of peripheral pathway represents an important advance towards fully understanding the whole 3-PBA metabolism in M-4. A specific altered metabolization was found for the first time, that is, resting cells of M-4 skipped the reduction step and initiate hydroxylation directly, by comparison with growing cells. Transcriptome analysis indicated that 3-PBA induced the up-regulation of genes related to energy investment, oxidative stress response, membrane transport and DNA repair. In-depth functional interpretation of differential expression genes suggested that the generation 3-PBOH and hydroxylated 3-PBA may be due to the participation of flavin-dependent monooxygenases (FMOs) and cytochrome P450 (CYP450), respectively. This study provides new insight to reveal the biodegradation mechanism of 3-PBA by A . oryzae M-4.
研究领域:
环境科学
·
微生物学
·
分子生物学
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LANGMUIR
·
2021
·
IF 3.882
Yixian Luo,Dou Jin,Wenjin He,Jianzhong Huang,Aicheng Chen,Feng Qi
The opal-like SiO2 microcarriers with different pore diameters named opal-SiO2I and opal-SiO2II were synthesized and utilized as microcarriers to immobilize Rhizopus oryzae lipase (ROL) and Aspergillus oryzae α-amylases (AOA). ROL and AOA can be more stably immobilized on the cross-linked SiO2 opals by neopentyl glycol diglycidyl ether (NGDE), which is the first attempt to use it as a cross-linking agent compared with glutaraldehyde. According to the morphology analysis, multiple layers of SiO2 monodisperse microspheres were regularly packed and formed an opal-like structure, and enzymes were well scattered and immobilized throughout the SiO2 opals. The results showed that the performance of enzymes immobilized on opal-SiO2II with a larger specific surface area was much better than that of opal-SiO2I. The enzyme activity of ROL@opal-SiO2II and AOA@opal-SiO2II cross-linked with 1% NGDE increased 5.32 and 9.32 times compared with their free counterpart, respectively. Furthermore, pH and thermal stability and reusability of ROL/AOA@opal-SiO2II were significantly improved and higher than those of ROL/AOA@opal-SiO2I and free enzymes. This study provides an easily obtained microcarrier opal-SiO2II, which shows potential for efficient different enzyme immobilizations and further industrial applications.
研究领域:
生物化学
·
材料科学
·
酶工程
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RENEWABLE ENERGY
·
2024
·
IF 9
Shuaibo Xia,Cai Shen,Jiale Lin,Maolin Tu,Chin-Ping Tan,Ling-Zhi Cheong
The absorption process of Aspergillus oryzae lipase (AOL) onto the surface of ZIF-8 was elucidated using molecular dynamic (MD) simulation. AOL are absorbed onto the surface of ZIF-8 via hydrogen bonding, electrostatic and van der Waals interaction with minimal changes to its native configurations. Unlike free AOL, AOL@ZIF-8 can maintain its structure in simulated methanol solutions. To verify the results from MD simulation, AOL was immobilized onto ZIF-8 (AOL@ZIF-8: immobilization efficiency of 64.71 % and a protein loading of 57.36 mg/g) and characterized using SEM, FTIR, and XRD. As compared to free AOL, AOL@ZIF-8 demonstrated higher transesterification activity with an activity recovery of 106.18 %. In agreement with MD simulations, AOL@ZIF-8 exhibited enhanced stability in wide range of pH and under presence of alcohol solution. AOL@ZIF-8 can be used to catalyze transesterification of used cooking oil for production of biodiesel (FAME content of 81.19 %).
研究领域:
生物技术
·
化学工程
·
生物化学
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高影响因子
Frontiers in Molecular Biosciences
·
2021
·
IF 5.246
Qu Tianle,Zhang Chunyue,Qin Zhen,Fan Liqiang,Jiang Lihua,Zhao Liming
Aminooligosaccharides possess various biological activities and can exploit wide applications in food, pharmaceutical and cosmetic industries. Commercial aminooligosaccharides are often prepared by the hydrolysis of chitin and chitosan. In this study, a novel GH family 20 β-N-acetylhexosaminidases gene named AoNagase was cloned from Aspergillus oryzae and expressed in Pichia pastoris. The purified AoNagase had maximal activity at pH 5.5 and 65°C. It exhibited good pH stability in the range of pH 6.0–7.5 and at temperatures below 50°C. AoNagase was capable of hydrolyzing not only colloidal chitosan (508.26 U/mg) but also chitin (29.78 U/mg). The kinetic parameters (Km and Vmax) of AoNagase were 1.51 mM, 1106.02 U/mg for chitosan and 0.41 mM, 40.31 U/mg for colloidal chitin. To our knowledge, AoNagase is the first GH family 20 β-N-acetylhexosaminidase capable of hydrolyzing both chitosan and chitin. AoNagase is an endo-type β-N-acetylhexosaminidases and can potentially be used for the manufacturing of aminooligosaccharides.
研究领域:
生物化学
·
酶学
·
生物技术
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Frontiers in Microbiology
·
2023
·
IF 5.2
Li Heng,Lu Zhen-Ming,Deng Wei-Qin,Zhang Qi-Sheng,Chen Gong,Li Qi,Xu Zheng-Hong,Ma Yan-He
Broad bean paste-meju was fermented by a mixture of broad bean koji and saline; koji fermentation is an essential process for the production of broad bean paste-meju. Aspergillus oryzae was the most widely used in sauce fermentation. The purpose of this study was to research the factory adaptability of the highly efficient A. oryzae PNM003 and further evaluate the effect of fermentation conditions and fermentation strains on koji. A. oryzae PNM003 was compared with the widely used strain HN 3.042 not only in the laboratory but also in factory conditions (large scale). Results showed that the koji made with the same starter in the factory had a greater amount of fungi than that in the laboratory. Bacteria and yeast levels in HN_L koji were higher than in PN_L koji. As for fungi constitution, almost only Aspergillus survived in the end through the microorganism self-purification process during koji fermentation. As for the bacterial constitution, koji was grouped by fermentation conditions instead of fermentation starter. PN koji had higher protease activity and a higher content of total acids, amino acid nitrogen, amino acids, and organic acids in the laboratory conditions. Nevertheless, in factory conditions, PN koji and HN koji had similar indexes. As for volatile flavor compounds, koji made with the two starters in the same condition was grouped together. As for the same starter, there were more flavor compounds metabolized in the factory condition than in the laboratory condition, especially esters and alcohols. The results showed PN was a highly efficient strain to ferment koji, but the advantages were expressed more remarkably in laboratory conditions. In brief, the fermented condition had a greater influence than the fermentation starter for broad bean koji.
研究领域:
食品科学
·
微生物学
·
发酵技术
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2020
·
IF 4.192
Xianli Gao,Ermeng Liu,Yiyun Yin,Lixin Yang,Qingrong Huang,Sui Chen,Chi-Tang Ho
Aspergillus oryzae 3.042 was mutagenized using atmospheric and room-temperature plasma (ARTP) technology to enhance its salt-tolerant proteases activity. Compared to the starting strain, mutant H8 subjected to 180 s of ARTP treatment exhibited excellent genetic stability (15 generations), growth rate, and significantly increased activities of neutral proteases, alkaline proteases, and aspartyl aminopeptidase during fermentation. Mutant H8 significantly enhanced the contents of 1–5 kDa peptides, aspartic acid, serine, threonine, and cysteine in soy sauce by 16.61, 7.69, 17.30, 8.61, and 45.00%, respectively, but it had no effects on the contents of the other 14 free amino acids (FAAs) due to its slightly enhanced acidic proteases activity. Analyses of transcriptional expressions of salt-tolerant alkaline protease gene (AP, gi: 217809) and aspartyl aminopeptidase gene (AAP, gi: 6165646) indicated that their expression levels were increased by approximately 30 and 27%, respectively. But no mutation was found in the sequences of AP and AAP expression cassettes, suggesting that the increased activities of proteases in mutant H8 should be partially attributed to the increased expression of proteases. ARTP technology showed great potential in enhancing the activities of salt-tolerant proteases from A. oryzae.
研究领域:
微生物学
·
生物技术
·
食品科学
·
遗传学
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高影响因子
危险品化学品经营许可证（带存储）