Study on enzymatic activity and lipase catalysis by lipase high-yield strain

Abstract Background Prairie grass (Bromus catharticus) is a typical cool-season forage crop with high biomass production and fast growth rate during winter and spring. However, its genetic research and breeding has remained stagnant due to limited available genomic resources. The aim of this study was to generate large-scale genomic data using high-throughput transcriptome sequencing, and perform a preliminary validation of EST-SSR markers of B. catharticus. Results Eleven tissue samples including seeds, leaves, and stems were collected from a new high-yield strain of prairie grass BCS1103. A total of 257,773 unigenes were obtained, of which 193,082 (74.90%) were annotated. Comparison analysis between tissues identified 1803, 3030, and 1570 genes specifically and highly expressed in seed, leaf, and stem, respectively. A total of 37,288 EST-SSRs were identified from unigene sequences, and more than 80,000 primer pairs were designed. We synthesized 420 primer pairs and selected 52 ones with high polymorphisms to estimate genetic diversity and population structure in 24 B. catharticus accessions worldwide. Despite low diversity indicated by an average genetic distance of 0.364, the accessions from South America and Asia and wild accessions showed higher genetic diversity. Moreover, South American accessions showed a pure ancestry, while Asian accessions demonstrated mixed internal relationships, which indicated a different probability of gene flow. Phylogenetic analysis clustered the studied accessions into four clades, being consistent with phenotypic clustering results. Finally, Mantel analysis suggested the total phenotypic variation was mostly contributed by genetic component. Stem diameter, plant height, leaf width, and biomass yield were significantly correlated with genetic data (r > 0.6, P < 0.001), and might be used in the future selection and breeding. Conclusion A genomic resource was generated that could benefit genetic and taxonomic studies, as well as molecular breeding for B. catharticus and its relatives in the future.

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Optimization of the Fermentation Conditions for 1-Deoxynojirimycin Production by Streptomyces lawendulae Applying the Response Surface Methodology

International Journal of Food Engineering ◽

10.2202/1556-3758.2354 ◽

2011 ◽

Vol 7 (3) ◽

Cited By ~ 14

Author(s):

Zhao-Jun Wei ◽

Le-Chun Zhou ◽

Hua Chen ◽

Gui-Hai Chen

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Starch Content ◽

Ultra Violet ◽

Soluble Starch ◽

High Yield ◽

Yield Strain ◽

Fermentation Time ◽

Ethyl Sulfate ◽

Initial Ph

Moranoline (1-Deoxynojirimycin, DNJ) is a piperidine alkaloid, and shows high inhibit activities to glucoamylase and ?-glucosidase. One DNJ high-yield strain of Streptomyces lawendulae was obtained after isolated form soil and mutated with the ultra violet (UV) and ethyl sulfate (DES), which named as TB-412, and can produce DNJ with 35.925 mg/L. Response surface methodology (RSM) was applied to optimize the parameters of DNJ yield from S. lawendulae TB-412. The effects of independent variables of fermentation, including time, temperature, initial pH and the soluble starch content were investigated. The statistical analysis showed that the fermentation time, pH and the soluble starch content, and the quadratics of time, temperature, pH and the soluble starch content, as well as the interactions between fermentation time and pH, and time and the soluble starch content, showed significant effects on DNJ yield. The optimal process parameters for DNJ production within the experimental range of the variables researched was at 11d, 27 °C, pH 7.5, and 8% soluble starch content. At this condition, the DNJ yield was predicted to be 42.875 mg/L.

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Uncovering and Engineering a Mini-Regulatory Network of the TetR-Family Regulator SACE_0303 for Yield Improvement of Erythromycin in Saccharopolyspora erythraea

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.692901 ◽

2021 ◽

Vol 9 ◽

Author(s):

Ying Liu ◽

Sabir Khan ◽

Panpan Wu ◽

Bowen Li ◽

Lanlan Liu ◽

...

Keyword(s):

Regulatory Network ◽

Target Genes ◽

Antibacterial Activities ◽

Saccharopolyspora Erythraea ◽

High Yield ◽

Yield Strain ◽

Yield Improvement ◽

Erythromycin Production

Erythromycins produced by Saccharopolyspora erythraea have broad-spectrum antibacterial activities. Recently, several TetR-family transcriptional regulators (TFRs) were identified to control erythromycin production by multiplex control modes; however, their regulatory network remains poorly understood. In this study, we report a novel TFR, SACE_0303, positively correlated with erythromycin production in Sac. erythraea. It directly represses its adjacent gene SACE_0304 encoding a MarR-family regulator and indirectly stimulates the erythromycin biosynthetic gene eryAI and resistance gene ermE. SACE_0304 negatively regulates erythromycin biosynthesis by directly inhibiting SACE_0303 as well as eryAI and indirectly repressing ermE. Then, the SACE_0303 binding site within the SACE_0303-SACE_0304 intergenic region was defined. Through genome scanning combined with in vivo and in vitro experiments, three additional SACE_0303 target genes (SACE_2467 encoding cation-transporting ATPase, SACE_3156 encoding a large transcriptional regulator, SACE_5222 encoding α-ketoglutarate permease) were identified and proved to negatively affect erythromycin production. Finally, by coupling CRISPRi-based repression of those three targets with SACE_0304 deletion and SACE_0303 overexpression, we performed stepwise engineering of the SACE_0303-mediated mini-regulatory network in a high-yield strain, resulting in enhanced erythromycin production by 67%. In conclusion, the present study uncovered the regulatory network of a novel TFR for control of erythromycin production and provides a multiplex tactic to facilitate the engineering of industrial actinomycetes for yield improvement of antibiotics.

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High-Throughput System for Screening of Cephalosporin C High-Yield Strain by 48-Deep-Well Microtiter Plates

Applied Biochemistry and Biotechnology ◽

10.1007/s12010-013-0095-4 ◽

2013 ◽

Vol 169 (5) ◽

pp. 1683-1695 ◽

Cited By ~ 22

Author(s):

Jun Tan ◽

Ju Chu ◽

Yuyou Hao ◽

Yuanxin Guo ◽

Yingping Zhuang ◽

...

Keyword(s):

High Throughput ◽

High Yield ◽

Cephalosporin C ◽

Yield Strain ◽

Deep Well ◽

Microtiter Plates

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A histological basis for differences in breast meat yield Between two strains of white turkeys

The Journal of Agricultural Science ◽

10.1017/s0021859600028999 ◽

1980 ◽

Vol 94 (2) ◽

pp. 383-388 ◽

Cited By ~ 2

Author(s):

H. J. Swatland

Keyword(s):

Growth Curves ◽

Live Weight ◽

Optimal Growth ◽

High Yield ◽

Muscle Fibres ◽

Breast Meat ◽

Yield Strain ◽

Breast Meat Yield ◽

Genetic Strains ◽

Similar Growth

SummaryTwo genetic strains of white turkeys were reared together in a large flock under conditions allowing optimal growth from hatching to 20 weeks. Birds of both strains followed similar growth curves with males of both strains growing heavier than females. In one strain, however, the yield of breast meat as a proportion of live weight was greater in both males (287 v. 225 g/kg) and females (276 v. 201 g/kg). When expressed in ratio to breast width, maximum meat depth was greater in the strain which yielded the higher proportion of breast meat. However, with this index of breast plumpness, females scored higher than males in both strains (high yield, 0·45:1·00 v. 0·37:1·00; low yield, 0·33:1·00 v. 0·31:1·00). In the superficial anterior region of the pectoralis muscle in both strains and sexes, there were strong adenosine triphosphatase and weak succinate dehydrogenase reactions in nearly all muscle fibres. Differences in the radial growth of muscle fibres were found between the two strains and, by 20 weeks, birds of the lower yield strain had reached only 0·85 (males) or 0·82 (females) of the mean diameters attained in birds of the higher yielding strain.

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An injectable meta-biomaterial

10.1101/2020.01.30.926931 ◽

2020 ◽

Author(s):

A. Béduer ◽

F. Bonini ◽

C. Verheyen ◽

P. Burch ◽

T. Braschler

Keyword(s):

Pore Space ◽

Tissue Mechanics ◽

High Yield ◽

Physical Parameters ◽

Yield Strain ◽

Dynamic Matching ◽

Tissue Integration ◽

One Year ◽

Elastic Softening

AbstractWe present a novel type of injectable biomaterial with an elastic softening transition. The material enables in-vivo shaping, followed by induction of 3D stable vascularized tissue adopting the desired shape. We establish the necessary geometrical and physical parameters by extensive numerical simulation. Irregular particle shape dramatically enhances yield strain for in-vivo stability against deformation, while friction and porosity provide the elastic softening transition as an emergent meta-material property. Accordingly, we synthesize our injectable meta-biomaterial as a suspension of irregularly fragmented, highly porous sponge-like microgels. The meta-biomaterial exhibits both high yield strain, and the desired novel elastic softening transition for in-situ shaping and unprecedented dynamic matching of adipose tissue mechanics. In vivo, predetermined shapes can be sculpted manually after subcutaneous injection in mice. The 3D shape is maintained during excellent host tissue integration into the particle pore space. The meta-biomaterial sustains vascularized connective tissue to the end of one-year follow-up.

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Breeding of a high-yield strain for commercial cultivation by crossing Pholiota adiposa and P. limonella

10.1101/728626 ◽

2019 ◽

Author(s):

Chengbo Rong ◽

Shuang Song ◽

Li Yang ◽

Jiachan Zhang ◽

Yurong Niu ◽

...

Keyword(s):

Ligninolytic Enzymes ◽

Edible Mushroom ◽

Inter Simple Sequence Repeat ◽

High Yield ◽

Yield Strain ◽

Hybrid Strain ◽

Commercial Cultivation ◽

Pcr Products ◽

Medicinal Properties ◽

Colonization Time

AbstractPholiota adiposa is an edible mushroom with excellent nutritional and medicinal properties. However, fruiting body yields are low, and the commercial cultivation potential of this fungus is limited. In the present study, 279 crossbred strains were obtained by mono-mono crossing of monokaryotic strains derived from P. adiposa HS5 and P. limonella HS4. Ligninolytic enzymes and mycelial growth rate were used as markers to screen the crossbred strains, and 18 were selected for further analysis. Crossbred strain A10B4 displayed the highest yield, i.e., 165.91 ± 12.56 g per bag, which was 31.34 g and 74.48 g more than that of strains HS5 and HS4, respectively. The mycelial colonization time of A10B4 was 25.18 ± 1.33 days, which was 5.64 days shorter than that of HS5. A10B4 was characterized by inter-simple sequence repeat molecular markers and antagonism tests. Differences in PCR products from parental and crossbred strains were observed. Therefore, the newly developed hybrid strain A10B4, named P. adiposa-limonella HS54 and having a high yield and desirable traits, might be suitable for commercial cultivation.

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Positive regulation of MarR-type regulator slnO and improving salinomycin production of Streptomyces albus by multiple transcriptional regulations

Canadian Journal of Microbiology ◽

10.1139/cjm-2021-0342 ◽

2022 ◽

Author(s):

Hongrui Zhang ◽

Weiwei Chen ◽

Xinyi Wang ◽

Yongquan Li ◽

Zhenhong Zhu

Keyword(s):

Gene Deletion ◽

High Yield ◽

Specific Gene ◽

Wild Type ◽

Yield Strain ◽

Positive Regulation ◽

Over Expression ◽

Streptomyces Albus ◽

Regulation Strategies ◽

Expression Strain

The purpose of this study is to explore the function of MarR-family regulator slnO. In addition, the high-yield strain of salinomycin was constructed by using combined regulation strategies. Firstly the slnO gene over-expression strain (GO) was constructed in Streptomyces albus. Compared to wild type (WT) strain，salinomycin production in GO strain was increased about 28%. Electrophoretic mobility gel shift assays (EMSAs) confirmed that SlnO protein can bind specifically to the intergenic region of slnN-slnO, slnQ-slnA1 and slnF-slnT. qRT-PCR experiments also showed that slnA1, slnF, and slnT1 were significantly up-regulated, while the expression level of the slnN gene was down-regulated in GO strain. Secondly, slnN gene deletion strain (slnNDM) was used as the starting strain, and the pathway specific gene slnR in salinomycin gene cluster was over expressed in slnNDM. The new strain was named ZJUS01. The yield of salinomycin in ZJUS01 strain was 25% and 56% higher than that in slnNDM strain and WT strain. Above results indicate that the slnO gene has a positive regulation effect on the biosynthesis of salinomycin. Meanwhile, the yield of salinomycin could be greatly increased by manipulating multiple transcriptional regulations.

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Construction of a novel MK-4 biosynthetic pathway in Pichia pastoris through heterologous expression of HsUBIAD1

Microbial Cell Factories ◽

10.1186/s12934-019-1215-9 ◽

2019 ◽

Vol 18 (1) ◽

Cited By ~ 1

Author(s):

Xiaowen Sun ◽

Hui Liu ◽

Peng Wang ◽

Li wang ◽

Wenfeng Ni ◽

...

Keyword(s):

Pichia Pastoris ◽

Heterologous Expression ◽

Biosynthetic Pathway ◽

Initial Conditions ◽

Expression System ◽

Value Added ◽

Theoretical Research ◽

High Yield ◽

Yield Strain ◽

Synthetic Pathway

Abstract Background With a variety of physiological and pharmacological functions, menaquinone is an essential prenylated product that can be endogenously converted from phylloquinone (VK1) or menadione (VK3) via the expression of Homo sapiens UBIAD1 (HsUBIAD1). The methylotrophic yeast, Pichia pastoris, is an attractive expression system that has been successfully applied to the efficient expression of heterologous proteins. However, the menaquinone biosynthetic pathway has not been discovered in P. pastoris. Results Firstly, we constructed a novel synthetic pathway in P. pastoris for the production of menaquinone-4 (MK-4) via heterologous expression of HsUBIAD1. Then, the glyceraldehyde-3-phosphate dehydrogenase constitutive promoter (PGAP) appeared to be mostsuitable for the expression of HsUBIAD1 for various reasons. By optimizing the expression conditions of HsUBIAD1, its yield increased by 4.37 times after incubation at pH 7.0 and 24 °C for 36 h, when compared with that under the initial conditions. We found HsUBIAD1 expressed in recombinant GGU-23 has the ability to catalyze the biosynthesis of MK-4 when using VK1 and VK3 as the isopentenyl acceptor. In addition, we constructed a ribosomal DNA (rDNA)-mediated multi-copy expression vector for the fusion expression of SaGGPPS and PpIDI, and the recombinant GGU-GrIG afforded higher MK-4 production, so that it was selected as the high-yield strain. Finally, the yield of MK-4 was maximized at 0.24 mg/g DCW by improving the GGPP supply when VK3 was the isopentenyl acceptor. Conclusions In this study, we constructed a novel synthetic pathway in P. pastoris for the biosynthesis of the high value-added prenylated product MK-4 through heterologous expression of HsUBIAD1 and strengthened accumulation of GGPP. This approach could be further developed and accomplished for the biosynthesis of other prenylated products, which has great significance for theoretical research and industrial application.

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Transcriptomic resources for prairie grass (Bromus catharticus): expressed transcripts, tissue-specific genes, and identification and validation of EST-SSR markers

10.21203/rs.3.rs-306875/v1 ◽

2021 ◽

Author(s):

Ming Sun ◽

Zhixiao Dong ◽

Jian Yang ◽

Wendan Wu ◽

Chenglin Zhang ◽

...

Keyword(s):

Genetic Diversity ◽

Ssr Markers ◽

Large Scale ◽

Genetic Research ◽

High Yield ◽

Yield Strain ◽

Tissue Samples ◽

Prairie Grass ◽

The Future ◽

Genomic Resource

Abstract Background: Prairie grass (Bromus catharticus) is a typical cool-season forage crop with high biomass production and fast growth rate during winter and spring. However, its genetic research and breeding has remained stagnant due to limited available genomic resources. The aim of this study was to generate large-scale genomic data using high-throughput transcriptome sequencing, and perform a preliminary validation of EST-SSR markers of B. catharticus.Results: Eleven tissue samples including seeds, leaves, and stems were collected from a new high-yield strain of prairie grass BCS1103. A total of 257,773 unigenes were obtained, of which 193,082 (74.90%) were annotated. Comparison analysis between tissues identified 1803, 3030, and 1570 genes specifically expressed in the seed, leaf, and stem, respectively. A total of 37,288 EST-SSRs were identified from unigene sequences, and more than 80,000 primer pairs were designed. We synthesized 420 primer pairs and selected 52 ones with high polymorphisms to estimate genetic diversity and population structure in 24 B. catharticus accessions worldwide. Despite low diversity indicated by an average genetic distance of 0.358, the accessions from South America and Asia and wild accessions showed higher genetic diversity. Moreover, South American accessions showed a pure ancestry, but Asian accessions demonstrated mixed relationships, which indicated a different probability of gene flow among the two regions. Phylogenetic analysis clustered the studied accessions into four clades. Finally, phenotypic clustering and Mantel analysis suggested the total phenotypic variation was mostly contributed by the genetic component. Stem diameter, plant height, leaf width, and biomass yield significantly correlated with genetic data (r > 0.6, P < 0.001), and might be genetically stable in the future selection and breeding.Conclusion: A genomic resource was generated that could benefit genetic and taxonomic studies, as well as molecular breeding for B. catharticus and it relatives in the future.

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