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Transcription analysis of hyaluronan biosynthesis genes in Streptococcus zooepidemicus and metabolically engineered Lactococcus lactis
Shashi Bala Prasad,
Published in
2012
PMID: 22367612
Volume: 94
   
Issue: 6
Pages: 1593 - 1607
Abstract
The has operon genes in the hyaluronan (HA) producer, Streptococcus zooepidemicus, encode for some of the critical enzymes in the HA biosynthetic pathway. Heterologous expression of different combinations of multiple has genes has resulted in increasing HA production to varying degrees in different recombinant strains. In this work, a recombinant Lactococcus lactis strain (SJR6) was constructed, with insertion of three has operon genes (hasABD) from S. zooepidemicus. The SJR6 strain was found to be a better HA producer than two previously constructed recombinant L. lactis strains (SJR2 and SJR3), containing hasAB and hasABC genes, respectively, but exhibited lower HA production than the native HA producer S. zooepidemicus. To understand the differences in HA yield between the various strains, transcriptions of the HA biosynthesis genes (has genes and their homologues) were compared at different phases of exponential growth of the L. lactis and S. zooepidemicus cultures. The mRNA levels of all the heterologous has genes were expectedly far higher than their corresponding homologues in the L. lactis strains. The relative mRNA level of the hasB-homologue, viz. ugd (encoding UDP-glucose dehydrogenase), was found to be much lower than that of other homologues, corroborating earlier reports which indicate tight transcriptional regulation of the ugd gene in L. lactis. Interestingly, all the has gene homologues were found to be up-regulated in all the recombinant L. lactis strains, when compared with the corresponding genes in the untransformed strain, L. lactis NZ9000. A transcription analysis of S. zooepidemicus cultures revealed that the has operon was down-regulated in the midexponential growth phase in comparison to the early- and lateexponential growth phases. The transcription analyses in this study have provided insights for the design of recombinant strains with higher HA productivity. © Springer-Verlag 2012.
About the journal
JournalApplied Microbiology and Biotechnology
ISSN01757598
Open AccessNo
Concepts (53)
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    HYALURONAN
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    LACTOCOCCUS LACTIS
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    REAL-TIME PCR
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    STREPTOCOCCUS ZOOEPIDEMICUS
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    TRANSCRIPTION ANALYSIS
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    Biochemistry
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    Biosynthesis
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    Encoding (symbols)
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    Gene encoding
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    Hyaluronic acid
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    Metabolic engineering
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    Polymerase chain reaction
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    Strain
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    Transcription
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    Genomic dna
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    Bacterial protein
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    URIDINE DIPHOSPHATE GLUCOSE DEHYDROGENASE
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    Bacterium
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    Bioengineering
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    Enzyme activity
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    GROWTH MODELING
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    Metabolism
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    Real time
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    Recombination
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    Article
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    Bacillus subtilis
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    Bacterial cell wall
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    Bacterial gene
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    Bacterial growth
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    Bacterial strain
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    Bacterium culture
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    Biomass production
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    Controlled study
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    Down regulation
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    Gene expression
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    Genetic transcription
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    LACTOCOCCUS LACTIS
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    Microbial biomass
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    Nonhuman
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    Real time polymerase chain reaction
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    RNA ANALYSIS
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    STREPTOCOCCUS EQUI ZOOEPIDEMICUS
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    Enzymology
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    Gene expression regulation
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    Genetics
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    STREPTOCOCCUS EQUI
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    Bacterial proteins
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    Biosynthetic pathways
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    Gene expression regulation, bacterial
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    LACTOCOCCUS LACTIS
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    STREPTOCOCCUS EQUI
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    Transcription, genetic
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    URIDINE DIPHOSPHATE GLUCOSE DEHYDROGENASE