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酱香型白酒第2、3轮次堆积发酵有机酸代谢规律与菌群结构及其相关性解析

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食品科学 2024年第18期45卷 106-115页

作者：黄廷财柴丽娟时伟吴双全杨波张晓娟陆震鸣王松涛沈才洪许正宏江南大学生物工程学院工业生物技术教育部重点实验室江苏无锡214122 江南大学粮食发酵与食品生物制造国家工程研究中心江苏无锡214122 贵州中鉴酒业集团有限公司贵州遵义563000 国家固态酿造工程技术研究中心四川泸州646000

本实验以酱香型白酒优势产酒轮次(即第3、4轮次基酒所对应的第2、3轮次)堆积发酵过程中的酒醅为研究对象,采用高效液相色谱和高通量测序等技术对酒醅中的有机酸及菌群结构进行分析,探讨堆积发酵过程中有机酸与微生物群落之间的互作关系... 详细信息

本实验以酱香型白酒优势产酒轮次(即第3、4轮次基酒所对应的第2、3轮次)堆积发酵过程中的酒醅为研究对象,采用高效液相色谱和高通量测序等技术对酒醅中的有机酸及菌群结构进行分析,探讨堆积发酵过程中有机酸与微生物群落之间的互作关系。结果表明,堆积发酵酒醅中主要有机酸为乳酸(32.70 mg/g,醅样内平均含量,后同)、乙酸(4.87 mg/g)和琥珀酸(1.93 mg/g)。第2、3轮次堆积发酵酒醅中共有8个优势细菌属和7个优势真菌属(平均相对丰度>1%),细菌的优势菌属主要为枝芽孢杆菌属(Virgibacillus)、克罗彭斯特菌属(Kroppenstedtia)和芽孢杆菌属(Bacillus);真菌的优势菌属主要为红曲霉属(Monascus)、嗜热真菌属(Thermomyces)和嗜热子囊菌属(Thermoascus)。主坐标分析表明,第2、3轮次堆积发酵细菌群落结构存在显著差异,真菌差异不显著。共现性网络分析表明,细菌-真菌(55.60%)、细菌-细菌(50.57%)和真菌-真菌(91.67%)网络均以正相关为主,且细菌-细菌群落之间存在更多的未知互作关系。Mantel test分析表明,酸度与细菌群落结构呈极显著正相关(Plus)、芽孢杆菌属、鞘氨醇杆菌属(Sphingobacterium)呈正相关,与高温放线菌属(Thermoactinomyces)呈负相关。研究结果可为优质酱酒的生产调控提供一些理论基础。

关键词：酱香型白酒堆积发酵微生物群落结构有机酸

酱香型白酒堆积酒醅中象牙色克罗彭斯特德菌的分离筛选及其代谢特性解析

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微生物学报 2024年第7期64卷 2502-2521页

作者：吴双全柴丽娟黄廷财时伟陆震鸣张晓娟王松涛沈才洪许正宏江南大学生物工程学院江苏无锡214122 江南大学粮食发酵与食品生物制造国家工程研究中心江苏无锡214122 国家固态酿造工程技术研究中心四川泸州646000

【目的】高温堆积发酵是酱香型白酒生产中的关键工艺环节,克罗彭斯特德菌属(Kroppenstedtia)是堆积酒醅中的优势细菌属,研究其生长和代谢特征对于理解堆积发酵的关键作用至关重要。【方法】采用胰酪大豆蛋白胨(tryptic soy broth,TSB)... 详细信息

【目的】高温堆积发酵是酱香型白酒生产中的关键工艺环节,克罗彭斯特德菌属(Kroppenstedtia)是堆积酒醅中的优势细菌属,研究其生长和代谢特征对于理解堆积发酵的关键作用至关重要。【方法】采用胰酪大豆蛋白胨(tryptic soy broth,TSB)培养基从酒醅中筛选克罗彭斯特德菌,通过形态学和16S rRNA基因测序确定其分类学地位,结合菌株纯培养及不同温度(45℃和50℃)下的高粱固态发酵实验,研究其生长和挥发性化合物代谢特征。【结果】从酱香型白酒堆积酒醅中分离筛选到3株克罗彭斯特德菌,经鉴定为象牙色克罗彭斯特德菌(Kroppenstedtia eburnea)。液态培养时菌株*** 1613促进吡嗪类物质的产生,为对照的2.66倍。在固态发酵高粱中检出的挥发性化合物主要为醇类和酸类,总含量随发酵时间的推移而逐渐增加。50℃发酵高粱有利于醇类、酸类和吡嗪类物质的积累,而45℃下酯类物质含量较高。3株菌以高粱为基质进行固态发酵时主要代谢产物是苯乙醇和异戊酸,其中*** 1615在50℃下发酵15 d时苯乙醇和异戊酸含量最高,为(31.17±0.14)μg/g和(16.75±0.76)μg/g。菌株*** 6E22在50℃下发酵15 d时2,5-二甲基吡嗪含量最高,为(1.67±0.14)μg/g。菌株*** 1613在发酵15 d时己酸含量最高,为(3.74±0.19)μg/g。50℃下发酵高粱自身中醛酮类物质积累明显。偏最小二乘判别分析(partial least squares discrimination analysis,PLS-DA)显示,温度和时间对3株菌发酵高粱中挥发性化合物的组成有显著影响。【结论】象牙色克罗彭斯特德菌有助于堆积发酵酒醅风味化合物的产生,特别是醇类、酸类和吡嗪类等酱香型白酒特征风味物质。

关键词：堆积发酵象牙色克罗彭斯特德菌固态发酵挥发性化合物

A Surfactant-stable Bacillus pumilus K9 α-Keratinase and Its Potential Application in Detergent Industry

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Chemical Research in Chinese Universities 2015年第1期31卷 91-97页

作者： GONG Jinsong WANG Yue ZHANG Dandan LI Heng ZHANG Xiaomei ZHANG Rongxian lu zhenming XU Zhenghong SHI Jinsong School of Pharmaceutical Science Jiangnan University Wuxi 214122 P. R. China

An a-keratinase producing strain was isolated with wool as the sole carbon and nitrogen source and identified as Bacilluspumilus K9. The major amino acids liberated from the keratin degradation of wool by B. pumilus K9 were glutamic acid and leucine. The a-keratinase was purified to electrophoretic homogeneity with a molecular weight of 32000. The purified enzyme exhibits an optimum activity at 60 ℃ and pH=9.0. It was stable at pH values between 8 and 11. Bacillas pumilus keratinase displays a high activity towards casein, keratin, wool and feather, which indicates its wide application range. The keratinase was completely inhibited by phenylmethyl-sulfonyl fluoride（PMSF） and β-mercaptoethanol, and moderately inhibited by ethylemediamine-tetraacetic acid（EDTA）, suggesting it is a metallo-cysteine keratinase. This enzyme could remain stable that could even be promoted in the presence of surfactants, including sodium dodecyl sulfate（SDS）, Tween and Triton. And Tween 40 and Triton X-100 could substantially enhance the activity of the enzyme by 54% and 35%, respectively. It may indicate the prominent feature of the keratinase to tolerate surfactants. The enzymatic properties distinguish this keratinase from others in the literature. Furthermore, this enzyme is extremely stable in the presence of a commercially available detergent with 1% concentration. Detergents ARIEL, Bluemoon and WhiteCat can enhance the activity of the keratinase by 43.56%, 15.22%, and 22.48%, respectively.

关键词： a-Keratinase Bacilluspumilus K9 Wool degradation Surfactant stability Detergent

基于宏基因组解析中温大曲成熟前后的微生物群落功能差异

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生物工程学报 2024年第3期40卷 877-894页

作者：刘文虎刘光钱张芮郑蕾陆震鸣张晓娟王松涛沈才洪史劲松许正宏柴丽娟江南大学粮食发酵与食品生物制造国家工程研究中心江苏无锡214122 江南大学生物工程学院江苏无锡214122 泸州品创科技有限公司四川泸州646000 国家固态酿造工程技术研究中心四川泸州646000 江南大学生命科学与健康工程学院江苏无锡214122

大曲是白酒酿造中的糖化发酵剂和生香剂,其成熟过程是获得高品质大曲的重要步骤。前期研究探索了大曲成熟前后的微生物群落组成和多样性,但对微生物群落功能的变化缺乏深入研究。本文在分析中温大曲成熟前后酶活和挥发性化合物的基础上... 详细信息

大曲是白酒酿造中的糖化发酵剂和生香剂,其成熟过程是获得高品质大曲的重要步骤。前期研究探索了大曲成熟前后的微生物群落组成和多样性,但对微生物群落功能的变化缺乏深入研究。本文在分析中温大曲成熟前后酶活和挥发性化合物的基础上,采用宏基因组学解析微生物群落组成和潜在功能的差异,并探索酶活和重要挥发性化合物变化的微生物基础。结果显示,大曲成熟后,水分含量(P≤0.05)、淀粉含量、液化力、糖化力(P≤0.05)和发酵力降低,而酸度和酯化力(P≤0.05)升高;同时,其挥发性化合物组成变化显著(P=0.001),芳香醇类和芳香酯类物质含量显著降低,吡嗪类、酮类和高级脂肪醇类物质含量显著升高。成熟大曲中毛霉菌目(Mucorales,34.8%−23.0%)和散囊菌目(Eurotiales,34.3%−20.1%)等真菌相对丰度降低,功能分析表明这降低了α-淀粉酶、α-葡萄糖苷酶、乙醇脱氢酶和乙醇脱氢酶(NADP+)的基因丰度(P>0.05),导致大曲液化力(P>0.05)、糖化力(P≤0.05)、发酵力(P>0.05)和苯乙醇等芳香醇类物质含量降低(P≤0.05)。此外,酵母菌目(Saccharomycetales,2.9%−16.6%)、乳杆菌目(Lactobacillales,14.9%−23.6%)和芽孢杆菌目(Bacillales,0.8%−3.8%)等微生物的相对丰度升高,使得乙酰基转移酶、羧酸酯酶、乙酰乳酸脱羧酶、R-乙偶姻脱氢酶和S-乙偶姻脱氢酶的基因丰度显著升高(P≤0.05),进而显著提高了大曲的酯化力和吡嗪类物质含量(P≤0.05)。本文在基因水平解析了大曲成熟前后酶活和重要挥发性化合物变化的微生物基础,可为理性调控大曲生产提供理论支撑。

关键词：中温大曲成熟宏基因组学微生物群落结构与功能

采用实时荧光定量PCR法检测大曲中的3种高温放线菌

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食品与发酵工业 2024年第18期50卷 300-308页

作者：陈政柴丽娟张晓娟许泓瑜史劲松王松涛张宿义沈才洪许正宏陆震鸣江南大学生物工程学院江苏无锡214122 江南大学粮食发酵与食品生物制造国家工程研究中心江苏无锡214122 江南大学生命科学与健康工程学院江苏无锡214122 国家固态酿造工程技术研究中心四川泸州646000 四川大学轻工科学与工程学院四川成都610065

建立实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)方法对大曲中3种高温放线菌(Thermoactinomyces vulgaris、***和***)进行特异性定量检测。根据***、***、***全基因组中特异性单拷贝核心基因recA、mdH、gyrA为参考序列分别设计了一对可扩增278、233、291 bp片段的引物;经特异性、有效性、准确性实验显示,在常见的13种白酒酿造微生物中,引物TV、TI、TD分别可特异性检测***、***、***,检测范围为2.82~8.82 lg copies/μL,加标回收率95%~105%。进一步对大曲发酵过程样品检测,结果显示,***含量在呈现波动趋势,在第5天达到最高,为(6.06±0.02)lg copies/g;***含量呈现先上升后下降的趋势,在第7天达到最高,为(6.33±0.13)lg copies/g;***含量在0~12 d呈现波动趋势,在12~14 d下降,随后一直上升,发酵结束时含量最高,为(7.62±0.02)lg copies/g。该研究建立的RT-qPCR方法可对大曲发酵过程中3种高温放线菌进行特异性鉴定和快速定量,为进一步监测高温放线菌在白酒酿造过程中的功能提供了方法。

关键词：大曲高温放线菌特异性引物实时荧光定量PCR

Multi-Component Resource Recycling from Waste Light-Emitting Diode Under Hydrothermal Condition:Plastic Package Degradation,Speciation of Nano-TiO_(2),and Environmental Impact Assessment

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Engineering 2024年第8期39卷 253-261页

作者： Yongliang Zhang lu Zhan zhenming Xu School of Environmental Science and Engineering Shanghai Jiao Tong UniversityShanghai 200240China

Light emitting diodes(LEDs)have accounted for most of the lighting market as the technology matures and costs continue to *** a new type of e-waste,LED is a double-edged sword,as it contains not only precious and rare metals but also organic packaging *** previous studies,LED recycling focused on recovering precious and strategic metals while ignoring harmful substances such as organic packaging *** crushing and other traditional methods,hydrothermal treatment can provide an environment-friendly process for decomposing packaging *** work developed a closed reaction vessel,where the degradation rate of plastic polyphthalamide(PPA)was close to 100%,with nano-TiO_(2)encapsulated in plastic PPA being efficiently recovered,while metals contained in LED were also recycled ***,the role of water in plastic PPA degradation that has been overlooked in current studies was explored and speculated in detail in this *** impact assessment revealed that the proposed recycling route for waste LED could significantly reduce the overall environmental impact compared to the currently published *** the developed method could reduce more than half the impact of global ***,this research provides a theoretical basis and a promising method for recycling other plastic-packaged e-waste devices,such as integrated circuits.

关键词： Waste LED Hydrothermal treatment Recycling Plastic PPA degradation Packaging materials

Molecular cloning and transcriptional analysis of a NPY receptor-like in common Chinese cuttlefish Sepiella japonica

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Journal of Oceanology and Limnology 2018年第3期36卷 892-904页

作者： YANG Jingwen XU Yuchao XU Ke PING Hongling SHI Huilai lu zhenming WU Changwen WANG Tianming National Engineering Research Center for Facilitated Marine Aquaculture Marine Science CollegeZhejiang Ocean UniversityZhoushan 316022China Marine Fisheries Research Institute of Zhejiang Province Zhoushan 316022China

Neuropeptide Y(NPY) has a pivotal role in the regulation of many physiological processes. In this study, the gene encoding a NPY receptor-like from the common Chinese cuttlefish Sepiella japonica( SjNPYR-like) was identified and characterized. The full-length SjNPYR-like cDNA was cloned containing a 492-bp of 5′ untranslated region(UTR), 1 182 bp open reading frame(ORF) encoding a protein of 393 amino acid residues, and 228 bp of 3′ UTR. The putative protein was predicted to have a molecular weight of 45.54 kDa and an isoelectric point(pI) of 8.13. By informatic analyses, SjNPYR-like was identified as belonging to the class A G protein coupled receptor(GPCR) family(the rhodopsin-type). The amino acid sequence contained 12 potential phosphorylation sites and five predicted N-linked glycosylation sites. Multiple sequence alignment and 3D structure modeling were conducted to clarify SjNPYR bioinformatics characteristics. Phylogenetic analysis identifies it as an NPYR with identity of 33% to Lymnaea stagnalis NPFR. Transmembrane properties of SjNPYR-like were demonstrated in vitro using HEK293 cells and the p EGFP-N1 plasmid. Relative quantifi cation of SjNPYR-like mRNA level confi rmed a high level expression and broad distribution of SjNPYR-like in various tissues of female S. japonica. In addition, the transcriptional profile of SjNPYR-like in the brain, liver, and ovary during gonadal development was analyzed. The results provide basic understanding on the molecular characteristics of SjNPYR-like and its potentially physical functions.

关键词： Sepiella japonica NPY receptor-like growth reproduction gene expression

Isolation via enrichment and characterization of ten polymorphic microsatellite loci in the cuttlefish, Sepiella maindroni de Rochebruns

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Acta Oceanologica Sinica 2010年第6期29卷 121-124页

作者： WU Changwen CHI Changfeng HE Guangyuan lu zhenming XU Meiying Key Laboratory of Molecular Biophysics Ministry of Education China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory Huazhong University of Science and Technology Wuhan 430074 China Marine Science School Zhejiang Ocean University Zhoushan 316004 China Zhejiang Dahaiyang Science and Technology CO. Ltd. Zhoushan 316004 China

Ten novel polymorphic microsatellite loci were developed using an enrichment and colony hybridization protocol and characterized for the cuttlefish,Sepiella *** was explored by using 30 individuals from the coastal waters of Zhanjiang,Guangdong province,China,in December *** number of alleles ranged from 5 to *** ranges of observed and expected heterozygosity were from 0.578 9 to 1.000 0 and 0.682 8 to 0.925 7,respectively,and the average polymorphic information content （P IC） was 0.778 *** microsatellite loci will certainly facilitate the detection of the genetic variation and population structure of ***.

关键词： SepieUa maindroni microsatellite DNA cuttlefish genetic variation

A Vector Tracking Loop Based on ML Estimation in Dynamic Weak Signal Environments

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A Vector Tracking Loop Based on ML Estimation in Dynamic Wea...