Promotion of spinosad biosynthesis by chromosomal integration of the Vitreoscilla hemoglobin gene in Saccharopolyspora spinosa

Luo, Ying; Kou, XiaoXiao; Ding, Xuezhi; Hu, Shengbiao; Tang, Ying; Li, Wenping; Fan, Huimin; Yang, Qi; Chen, HanNa; Xia, Liqiu

doi:10.1007/s11427-012-4276-0

Cited by 19 publications

(16 citation statements)

References 33 publications

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“…In the face of the obtained result regarding oxidoreductase activity in the NCYC3825 strain and the macroscopic characteristics of both, NCYC3825 and VHb+, strains, it can be postulated that modification of the red-ox potential mainly contributed to the observed differences between A18 and NCYC3825 strains. This statement is corroborated by the other literature data, where VHb gene was heterologously expressed and consequently lead to improvement of the modified strains’ performance during cultivation [ 54 - 57 ].…”

Section: Resultssupporting

confidence: 82%

A novel multigene expression construct for modification of glycerol metabolism in Yarrowia lipolytica

Celińska

Grajek

2013

Microb Cell Fact

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BackgroundHigh supply of raw, residual glycerol from biodiesel production plants promote the search for novel biotechnological methods of its utilization. In this study we attempted modification of glycerol catabolism in a nonconventional yeast species Yarrowia lipolytica through genetic engineering approach.ResultsTo address this, we developed a novel genetic construct which allows transferring three heterologous genes, encoding glycerol dehydratase, its reactivator and a wide-spectrum alcohol oxidoreductase under the control of glycerol-induced promoter. The three genes, tandemly arrayed in an expression cassette with a marker gene ura3, regulatory and targeting sequences (G3P dh promoter and XPR-like terminator, 28S rDNA as a target locus), were transferred into Yarrowia lipolytica cells. The obtained recombinant strain NCYC3825 was characterized at the molecular level and with respect to its biotechnological potential. Our experiments indicated that the novel recombinant strain stably borne one copy of the expression cassette and efficiently expressed heterologous alcohol oxidoreductase, while glycerol dehydratase and its reactivator were expressed at lower level. Comparative shake flask cultivations in glucose- and glycerol-based media demonstrated higher biomass production by the recombinant strain when glycerol was the main carbon source. During bioreactor (5 L) fed-batch cultivation in glycerol-based medium, the recombinant strain was characterized by relatively high biomass and lipids accumulation (up to 42 gDCW L-1, and a peak value of 38%LIPIDS of DCW, respectively), and production of high titers of citric acid (59 g L-1) and 2-phenylethanol (up to 1 g L-1 in shake flask cultivation), which are industrially attractive bioproducts.ConclusionsDue to heterogeneous nature of the observed alterations, we postulate that the main driving force of the modified phenotype was faster growth in glycerol-based media, triggered by modifications in the red-ox balance brought by the wide spectrum oxidoreductase. Our results demonstrate the potential multidirectional use of a novel Yarrowia lipolytica strain as a microbial cell factory.

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Section: Resultssupporting

confidence: 82%

A novel multigene expression construct for modification of glycerol metabolism in Yarrowia lipolytica

Celińska

Grajek

2013

Microb Cell Fact

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“…It may be possible to improve spinosad production if the oxygen supply is enhanced during high‐density fermentation. Luo et al found that expression of the Vitreoscilla hemoglobin gene can improve oxygen uptake and thus increase spinosad production. Since dissolved oxygen (DO) is an important influencing factor during the spinosad fermentation process, Bai et al established a four‐stage DO strategy for spinosad fermentation.…”

Section: Strategies For Spinosad Overproduction In Sa Spinosamentioning

confidence: 99%

Strategies for Enhancing the Yield of the Potent Insecticide Spinosad in Actinomycetes

Tao

Zhang

Deng

et al. 2018

Biotechnology Journal

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Spinosad is a potent insecticide that exhibits an excellent environmental and mammalian profile. However, spinosad production in the original producer, Saccharopolyspora spinosa, is insufficient for the huge global demand. Great efforts have been exerted to improve the production of spinosad. Strategies for spinosad overproduction in actinomycetes are reviewed in this article, including metabolic engineering of the precursor and spinosyn biosynthetic pathway, introduction of regulatory genes, genome-scale metabolic model-guided engineering, mutagenesis, genome shuffling, fermentation process optimization, omics analysis, and the heterologous biosynthesis of spinosad in other actinomycetes. Furthermore, highly productive industrial strains should be used as heterologous hosts for enhancing spinosad biosynthesis in the future. To accelerate the engineering process, the CRISPR/Cas9 system should be established in Sa. spinosa for large-scale genome editing. Notably, the regulatory mechanism of spinosad biosynthesis remains unclear. Thus, the combining multi-omics analysis with high-throughput screening of chemical elicitors would be a promising approach in characterizing the regulatory and signal transduction mechanisms and improving spinosad production in Sa. Spinosa.

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“…Vitreoscilla Hb is now being frequently exploited to increase the industrial production of proteins and other molecules by bacteria and yeasts under hypoxic reactor conditions [473–481], and the primary benefit appears to be increased microaerobic respiration vis-à-vis the NOD function [256, 258]. Another potential application is the use of the recently discovered dual function algal raphidophyte NOD-nitrate reductase [139] in bioremediation reactors to scavenge NO produced in fossil fuel combustion and to supply algae with nitrate for increased production of biofuels [482].…”

Section: Hb-nod Technologiesmentioning

confidence: 99%

Hemoglobin: A Nitric-Oxide Dioxygenase

Gardner

2012

Scientifica

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Members of the hemoglobin superfamily efficiently catalyze nitric-oxide dioxygenation, and when paired with native electron donors, function as NO dioxygenases (NODs). Indeed, the NOD function has emerged as a more common and ancient function than the well-known role in O2 transport-storage. Novel hemoglobins possessing a NOD function continue to be discovered in diverse life forms. Unique hemoglobin structures evolved, in part, for catalysis with different electron donors. The mechanism of NOD catalysis by representative single domain hemoglobins and multidomain flavohemoglobin occurs through a multistep mechanism involving O2 migration to the heme pocket, O2 binding-reduction, NO migration, radical-radical coupling, O-atom rearrangement, nitrate release, and heme iron re-reduction. Unraveling the physiological functions of multiple NODs with varying expression in organisms and the complexity of NO as both a poison and signaling molecule remain grand challenges for the NO field. NOD knockout organisms and cells expressing recombinant NODs are helping to advance our understanding of NO actions in microbial infection, plant senescence, cancer, mitochondrial function, iron metabolism, and tissue O2 homeostasis. NOD inhibitors are being pursued for therapeutic applications as antibiotics and antitumor agents. Transgenic NOD-expressing plants, fish, algae, and microbes are being developed for agriculture, aquaculture, and industry.

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Promotion of spinosad biosynthesis by chromosomal integration of the Vitreoscilla hemoglobin gene in Saccharopolyspora spinosa

Cited by 19 publications

References 33 publications

A novel multigene expression construct for modification of glycerol metabolism in Yarrowia lipolytica

A novel multigene expression construct for modification of glycerol metabolism in Yarrowia lipolytica

Strategies for Enhancing the Yield of the Potent Insecticide Spinosad in Actinomycetes

Hemoglobin: A Nitric-Oxide Dioxygenase

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