ENZYME AND MICROBIAL TECHNOLOGY
·
2021
·
IF 3.493
Miao Tian,Shaowei Huang,Zhiyuan Wang,Junying Fu,Pengmei Lv,Changlin Miao,Tao Liu,Lingmei Yang,Wen Luo
The propeptide is a short sequence that facilitates protein folding. In this study, four highly active Rhizomucor miehei lipase (RML) mutants were obtained through saturation mutagenesis at three propeptide positions: Ser8, Pro35, and Pro47. The enzyme activities of mutants P35 N, P47 G, P47 N, and S8E/P35S/P47A observed at 40 °C, and pH 8.0 were 10.19, 7.53, 6.15, and 8.24 times of that wild-type RML, respectively. The S8E/P35S/P47A mutant showed good thermostability. After incubation at 40 °C for 1 h, 98.98 % of its initial activity remained, whereas wild-type RML retained only 78.76 %. This result indicated that the enhancement of hydrophilicity of 35- and 47- amino-acid residues could promote the interaction between the propeptide and the mature peptide and the enzyme activity and expression level. Highly conserved sites had a more significant impact on enzyme performance than did other sites, similar to the Pro35 and Pro47 mutants showed in this study. This study provides a new idea for protein modification: enzyme performance can be improved through propeptide regulation.
研究领域:
生物化学
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酶学
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蛋白质工程
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产品关联
DOI
PubMed
高影响因子
Catalysts
·
2018
·Vol. 8
·期 3
·96
·
IF 3.465
Miaad Adnan,Kai Li,Li Xu,Yunjun Yan
This study presents a one-step encapsulation method for synthesizing X-shaped zeolitic imidazolate frameworks (ZIF-8) and immobilizingRhizomucor mieheilipase (RML). We proved that the morphological structure of ZIF-8 had changed after immobilization with enhanced characterization using a field-emission scanning electron microscope, an energy-dispersive spectrometer, a transmission electron microscope, a Fourier transform infrared spectrometer, and powder X-ray diffraction. The surface area and pore size of the carrier were investigated before and after immobilization using Brunauer–Emmett–Teller and Barrett–Joyner–Halenda methods, respectively. RML@ZIF-8 exhibited high recovery activity of up to 2632%, representing a 26-fold increase in its free lipase. Encapsulated RML was used for biodiesel production from soybean oil in an isooctane system with a conversion yield of 95.6% under optimum conditions. The resulting reusability of the immobilized enzyme indicated no substantial decline in the conversion yield, which remained at 84.7% of the initial activity after 10 cycles. The stability and high performance of the immobilized enzyme are attributed to the harmony between RML and ZIF-8 based on the easy synthesis of ZIF-8 and the short time required to immobilize RML.Keywords:X-shape ZIF-8;Rhizomucor mieheilipase (RML);encapsulation technique;biodiesel yield;soybean oil
研究领域:
生物化学
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材料科学
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可再生能源
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
·
2023
·
IF 3.3
Jun Wang,Feilong Zhang,Chuanli Zeng,Xiaoli Xi,Ting Yao,Haiyan Yang,Yanli Ma
Summary Sufu, a traditional Chinese fermented soybean food, had its flavour and quality influenced by the specific strains and fermentation environment. To investigate how microbial community succession affects the volatile aroma substances of Mucor racemosus sufu during fermentation, the microbial community structure using high-throughput sequencing (HTS) technology was analysed. Additionally, volatile flavour compounds using solid-phase microextraction gas chromatography mass spectrometry (SPME-GC–MS) were detected. Throughout the pre-fermentation, salting, and post-fermentation stages, the changes in the microbial communities over time and space due to environmental variations were observed. The colony compositions at the phylum and genus levels were also analysed. The results revealed that Proteobacteria and Actinobacteria were the dominant bacterial phyla in the final fermented product. Furthermore, several microorganisms associated with flavour substances were identified, such as Lactobacillus , Pseudomonas , Wescheria , Hyphomyces , Candida , and Pichia . These microorganisms primarily influenced the synthesis of alcohols and acids, contributing to the overall flavour profile of the sufu. In the analysis of the final product, it identified fifty-four volatile flavour compounds, including alcohols, esters, phenolic acids, aldehydes, ketones, ether, alkenes, and other compounds. These findings have important implications for isolating specific strains, enhancing flavour, and providing a solid foundation for optimising and ensuring the quality of sufu.
研究领域:
食品科学
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微生物学
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DOI
高影响因子
INDUSTRIAL CROPS AND PRODUCTS
·
2021
·
IF 5.645
Rongfei Zhang,Yingjun Cui,Meng Cheng,Yanli Guo,Xiangyou Wang,Juan Wang
Cinnamon essential oil-mesoporous silica nanoparticles (CEO-MSNPs)/potato starch films have better antifungal activity against molds commonly found in post-harvest white mushrooms. The antifungal effects of CEO-MSNPs against Mucor sp. FJ09 species and Mucor circinelloides CNRMA 03.0371 were investigated. The antifungal mechanisms of CEO-MSNPs against molds were elucidated. Results showed that after adding CEO-MSNPs at the minimum inhibitory concentration level, there were obvious changes in membrane permeability, reactive oxygen species (ROS) content, cell component leakage, cell component leakage, soluble sugar, and malondialdehyde (MDA) content. The scanning electron microscope assay showed that CEO-MSNPs effectively destroyed the mycelial morphology of Mucor sp. FJ09 species and Mucor circinelloides CNRMA 03.0371, and interfered with their normal growth. Notably, CEO-MSNPs could be in the up-regulation of nox1 and nox2 genes of Mucor sp. FJ09 species and Mucor circinelloides CNRMA 03.0371 to generate a large number of reactive oxygen species, thereby inhibiting spore germination and growth. The results of this study could provide a reference for screening key targets of drug resistance of molds in post-harvest white mushrooms.
研究领域:
食品科学
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微生物学
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纳米技术
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DOI
高影响因子
ENVIRONMENTAL RESEARCH
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2024
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IF 7.7
Xuan Zhu,Yi-Fan Zhao,Hong-Jian Wen,Yu Lu,Shuai You,Richard Ansah Herman,Jun Wang
Silkworm pupae, by-product of sericulture industry, is massively discarded. The degradation rate of silkworm pupae protein is critical to further employment, which reduces the impact of waste on the environment. Herein, magnetic Janus mesoporous silica nanoparticles immobilized proteinase K mutant T206M and Mucor circinelloides aspartic protease were employed in the co-degradation. The thermostability of T206M improved by enhancing structural rigidity ( t 1/2 by 30 min and T 50 by 5 °C), prompting the degradation efficiency. At 65 °C and pH 7, degradation rate reached the highest of 61.7%, which improved by 26% compared with single free protease degradation. Besides, the immobilized protease is easy to separate and reuse, which maintains 50% activity after 10 recycles. Therefore, immobilized protease co-degradation was first applied to the development and utilization of silkworm pupae resulting in the release of promising antioxidant properties and reduces the environmental impact by utilizing a natural and renewable resource.
研究领域:
生物技术
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环境科学
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蛋白质工程
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
·
IF 8.5
Xiang Zhou,Jiahao Xu,Jiayao Sun,Hao Chen,Dechun Huang,Wei Chen,Hongliang Qian
1,3-Dioleoyl-2-palmitoylglycerol (OPO) is crucial for infant nutrition; however, conventional immobilized lipase requires high-purity enzymes, which increases costs and limits industrial scalability. Herein, Rhizomucor miehei lipase (RML) was immobilized on surface-modified magnetic nanoparticles using cross-linked enzyme aggregates (CLEAs) technology to produce Fe 3 O 4 @SiO 2 @TPOAC@RML CLEAs. This approach combines the separation and immobilization of enzymes, allowing for the use of lower-purity lipase, which enhances its suitability for industrial-scale processes. The optimized Fe 3 O 4 @SiO 2 @TPOAC@RML CLEAs exhibited excellent thermal, pH, and storage stability. The performance of Fe 3 O 4 @SiO 2 @TPOAC@RML CLEAs was evaluated by catalyzing the synthesis of OPO through the enzymatic hydrolysis of glycerol tripalmitate (PPP) and oleic acid (OA). Under optimal conditions, the OPO content in products reached 63.33 ± 0.30%, highlighting the high efficiency of the enzymatic process. After eight reuse cycles, the Fe 3 O 4 @SiO 2 @TPOAC@RML CLEAs retained around 75% of their relative activity. Therefore, this strategy of lipase immobilization using CLEA technology, coupled with magnetic nanoparticles, offers a promising approach for OPO synthesis.
研究领域:
生物催化
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纳米技术
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酶固定化
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食品化学
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工业生物技术
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产品关联
DOI
PubMed
高影响因子
Journal of Fungi
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2022
·Vol. 8
·期 2
·119
·
IF 5.724
Larissa Balabanova,Aleksandra Seitkalieva,Yulia Yugay,Tatiana Rusapetova,Lubov Slepchenko,Anna Podvolotskaya,Margarita Yatsunskaya,Elena Vasyutkina,Oksana Son,Liudmila Tekutyeva,Yury Shkryl
An efficientAgrobacterium-mediated genetic transformation based on the plant binary vector pPZP-RCS2 was carried out for the multiple heterologous protein production in filamentous fungusThermothelomycesthermophilusF-859 (formerlyMyceliophthorathermophilaF-859). The engineered fungusTh.thermophiluswas able to produce plant storage proteins ofZeamays(α-zein Z19) andAmaranthushypochondriacus(albumin A1) to enrich fungal biomass by valuable nutritional proteins and improved amino acid content. The mRNA levels ofz19anda1genes were significantly dependent on their driving promoters: the promoter of tryptophan synthase (PtrpC) was more efficient to expressa1, while the promoter of translation elongation factor (Ptef) provided much higher levels ofz19transcript abundance. In general, the total recombinant proteins and amino acid contents were higher in the Ptef-containing clones. This work describes a new strategy to improve mycoprotein nutritive value by overexpression of plant storage proteins.Keywords:filamentous fungi;Myceliophthorathermophila;mycoprotein;recombinant protein;maize alpha-zein;amaranth albumin A1
研究领域:
微生物学
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分子生物学
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生物技术
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食品科学
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DOI
PubMed
高影响因子
Food Biophysics
·
2022
·Vol. 17
·期 4
·495-507
·
IF 3.34
Ghide Michael Kidane,Li Kai,Wang Jianhua,Abdulmalek Saadiah A.,Yan Yunjun
1,3-Dioleoyl-2-palmitoyl glycerol (OPO)-supplemented infant formulas mimic the nutritional properties of human milk, and thus are the best nutritional alternatives to breast feeding. Herein, structured triacyl glycerides rich in sn-1,3 oleic acid (OA) and sn-2 palmitic acid (PA) are synthesized via a new strategy by employing Rhizomucor miehei lipase (RML) adsorbed on magnetized multiwalled carbon nanotubes (mMWCNT) and synthesizing OPO-rich fats in a two-step enzymatic acidolysis. The results showed that the immobilized lipase (mMWCNT-COOH-RML) exhibited improved pH and thermal stability, and it could be separated conveniently from the reaction system via an external magnetic field. It also possessed a much better performance in catalysis of tripalmitin (PPP) with OA to synthesize OPO-rich fats in a two-step enzymatic acidolysis. Under optimized conditions, the content of palmitic acid at the sn-2 position and incorporation of OA at the sn-1,3 positions respectively reached 92.93% and 57.82%. mMWCNT-COOH-RML also exhibited better reusability and catalytic activity than the conventionally used commercial Lipozyme RM IM. The results indicate the employed new strategy for synthesis of OPO-rich fats has a prosperous potential in the application of infant formula formulations.
研究领域:
食品科学
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化学工程
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生物技术
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产品关联
DOI
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
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2023
·
IF 6
Yongyi Yang,Juan Peng,Qingqing Li,Qinfei Song,Quentin Cronk,Biao Xiong
Dark hawk tea, produced through pile-fermentation of Lauraceae family leaves, which is rich in various beneficial ingredients and health functions, relies heavily on microbial activity for its unique characteristics. However, the processing technology of dark hawk tea and the microorganisms' dynamic changes during its pile-fermentation remain largely unknown. In this study, based on the previous work, we optimized pile-fermentation parameters for analyzing changes in dark hawk tea's chemical components, volatile substances, and microbial diversity. The tea exhibited superior sensory scores and biochemical qualities under optimal conditions (35 °C, 62% water, 24 h), measuring 5.7 mg/g amino acid content. Gas chromatography-mass spectrometry (GC-MS) identified 108 volatile compounds, notably terpenes, ketones, and esters. Microbial analysis revealed Bacteroides and Faecalibacterium as dominant bacteria and Cladosporium and Mortierella as dominant fungi. Cladosporium prevailed in early fermentation, shifting to Mortierella later. Predicted metabolic pathways highlighted microbial functional genes predominantly participated in carbohydrate, amino acid, and energy metabolism during the pile-fermentation. These results hold important implications for expanding the variety of hawk tea products and deepening our understanding of microbial community dynamics through the pile-fermentation of dark hawk tea.
研究领域:
食品科学
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微生物学
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生物化学
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植物科学
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产品关联
DOI
高影响因子
BIORESOURCE TECHNOLOGY
·
2022
·
IF 11.889
Miao Tian,Lingmei Yang,Pengmei Lv,Zhiyuan Wang,Junying Fu,Changlin Miao,Zhibing Li,Lianhua Li,Tao Liu,Wenyi Du,Wen Luo
Exploiting highly active and methanol-resistant lipase is of great significance for biodiesel production. A semi-rational directed evolution method combined with N-glycosylation is reported, and all mutants exhibiting higher catalytic activity and methanol tolerance than the wild type (WT). Mutant N267 retained 64% activity after incubation in 50% methanol for 8 h, which was 48% greater than that of WT. The catalytic activity of mutants N267 and N167 was 30- and 71- fold higher than that of WT. Molecular dynamics simulations of N267 showed that the formation of new strong hydrogen bonds between glycan and the protein stabilized the structure of lipase and improved its methanol tolerance. N267 achieved biodiesel yields of 99.33% (colza oil) and 81.70% (waste soybean oil) for 24 h, which was much higher than WT (51.6% for rapeseed oil and 44.73% for wasted soybean oil). The engineered ProRML mutant has high potential for commercial biodiesel production.
研究领域:
生物化学
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生物技术
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酶工程
·
可再生能源
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DOI
高影响因子
ACS Sustainable Chemistry & Engineering
·
2019
·
IF 6.97
Abdul Basit,Ting Miao,Junquan Liu,Jiaqi Wen,Lina Song,Fengzhen Zheng,Huiqiang Lou,Wei Jiang
Here, we report highly efficient degradation of xylan into xylose by a single multifunctional xylanolytic enzyme from the filamentous fungus Thermothelomyces thermophila (termed Ttxy43). Ttxy43 shows three different enzyme activities toward carbohydrates, β-xylosidase (80.8 U/mg), endoxylanase (105.42 U/mg), and α-l-arabinofuranosidase enzyme activities (15.81 U/mg). Analysis of the catalytic mode of action of Ttxy43 for birchwood-xylan (BWX) reveals that endoxylanase initially degrades xylan to unbranched xylooligosaccharides (XOSs) (xylobiose, xylotriose, xylotetraose) as intermediates, which are then quickly hydrolyzed into single xylose by β-xylosidase. Site-directed mutagenesis studies indicate that Ttxy43 residues Asp134 and Glu228 are essential catalytic sites, while Glu176, Asp38, and Asp85 play an accessory role. More importantly, Ttxy43 displays higher degradation efficiency in comparison with a commercial β-xylosidase and endoxylanase “cocktail”. These findings elucidate an efficient integrated degradation mechanism of xylan under industrial reaction conditions, which provides a novel strategy to design techniques for biomass energy production.
研究领域:
生物化学
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生物技术
·
酶学
·
可再生能源
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产品关联
DOI
高影响因子
FOOD CHEMISTRY
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2022
·
IF 9.231
Lihong He,Can Zeng,Lingfeng Wei,Li Xu,Fenglin Song,Jianrong Huang,Nanjing Zhong
This study aims to fabricate immobilized lipases for efficient preparation of 1,3-dioleoyl-2-palmitoyl-glycerol (OPO) through acidolysis of glycerol tripalmitate (PPP). Twelve (three types) supports and five lipases were studied carefully. Among them, the immobilized Thermomyces lanuginosa lipase (TLL) samples exhibited overall better performance than that of other immobilized lipases. Particularly, organic groups functionalized SBA-15 (R-SBA-15) supported TLL (TLL@R-SBA-15) samples gave PPP conversion from 97.70 to 99.00 % and OPO content from 59.52 to 64.73 %. After optimization, PPP conversion up to 99.07 %, OPO content 73.15 % and sn -2 palmitic acid content 90.09 % were obtained with TLL@C 18 H 37 -SBA-15 as catalyst. Moreover, TLL@C 18 H 37 -SBA-15 exhibited better acidolysis performance from 50 °C than that from 60 to 80 °C, which helped inhibit acyl migration. In addition, after 5 cycles of reuse, TLL@C 18 H 37 -SBA-15 retained 81.04 % (based on OPO content) and 98.88 % (based on sn- 2 palmitic acid content) of its initial activity, indicating it had an attractive prospect in future applications.
研究领域:
生物化学
·
生物技术
·
催化
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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）