Expression of a synthetic Artemesia annua amorphadiene synthase in Aspergillus nidulans yields altered product distribution

Lubertozzi, David; Keasling, Jay D.

doi:10.1007/s10295-008-0400-3

Cited by 19 publications

(10 citation statements)

References 31 publications

(44 reference statements)

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“…This could be due to the low amounts produced in the plants or the sampling method used which could subject the major sesquiterpenes to chemically rearrange in different ways than in the in vitro studies, resulting in different compositions of minor sesquiterpene compounds. In addition, types of hosts may also influence composition of products [32]. There are also many other terpenes found in VMP open flowers especially monoterpenes which contribute to the fragrance of the plant.…”

Section: Resultsmentioning

confidence: 99%

Functional Expression of an Orchid Fragrance Gene in Lactococcus lactis

Song

Abdullah

et al. 2012

IJMS

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Vanda Mimi Palmer (VMP), an orchid hybrid of Vanda tesselata and Vanda Tan Chay Yan is a highly scented tropical orchid which blooms all year round. Previous studies revealed that VMP produces a variety of isoprenoid volatiles during daylight. Isoprenoids are well known to contribute significantly to the scent of most fragrant plants. They are a large group of secondary metabolites which may possess valuable characteristics such as flavor, fragrance and toxicity and are produced via two pathways, the mevalonate (MVA) pathway or/and the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway. In this study, a sesquiterpene synthase gene denoted VMPSTS, previously isolated from a floral cDNA library of VMP was cloned and expressed in Lactococcus lactis to characterize the functionality of the protein. L. lactis, a food grade bacterium which utilizes the mevalonate pathway for isoprenoid production was found to be a suitable host for the characterization of plant terpene synthases. Through recombinant expression of VMPSTS, it was revealed that VMPSTS produced multiple sesquiterpenes and germacrene D dominates its profile.

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Section: Resultsmentioning

confidence: 99%

Functional Expression of an Orchid Fragrance Gene in Lactococcus lactis

Song

Abdullah

et al. 2012

IJMS

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“…Artemisinic acid, a precursor of artemisinin, has been produced using Saccharomyces cerevisiae which engineered with FDS, ADS, and CYP71AV1 genes of A. annua . ADS gene which is a key enzyme in the cyclization of farnesyl pyrophosphate has also been expressed in Aspergilus nidulans [ 16 ] and Escherichia coli [ 17 ] to produce amorphadiene. However, these efforts have not shown a dramatic increase in the production of artemisinin and its precursor.…”

Section: Introductionmentioning

confidence: 99%

Optimization of genetic transformation of Artemisia annua L. Using Agrobacterium for Artemisinin production

2014

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Background:Artemisinin, a sesquiterpene lactone endoperoxide isolated from the medicinal plant Artemisia annua L., is a choice and effective drug for malaria treatment. Due to the low yield of artemisinin in plants, there is a need to enhance the production of artemisinin from A. annua and biotechnological technique may be one of the methods that can be used for the purpose.Aim:To study the transformation efficiency of Agrobacterium tumefaciens in A. annua that could be applied to enhance the production of artemisinin by means of transgenic plants.Setting and Designs:The factors influencing Agrobacterium-mediated transformation of A. annua were explored to optimize the transformation system, which included A. tumefaciens strain and effect of organosilicone surfactants. Three strains of A. tumefaciens, that is, LBA4404, GV1301, and AGL1 harboring the binary vector pCAMBIA 1303 have been used for transformation. The evaluation was based on transient β-glucuronidase (GUS).Materials and Methods:Plant cell cultures were inniatiated from the seeds of A. annua using the germination Murashige and Skoog medium. A. tumefaciens harboring pCAMBIA were tranformed into the leaves of A.annua cultures from 2-week-old-seedling and 2-month-old-seedling for 15 min by vacuum infiltration. Transformation efficiency was determinated by measuring of blue area (GUS expression) on the whole leaves explant using ImageJ 1.43 software. Two organosilicon surfactants, that is, Silwet L-77 and Silwet S-408 were used to improve the transformation efficiency.Results:The transformation frequency with AGL1 strain was higher than GV3101 and LBA4404 which were 70.91, 49.25, and 45.45%, respectively. Effect of organosilicone surfactants, that is, Silwet L-77 and Silwet S-408 were tested on A. tumefaciens AGL1 and GV3101 for their level of transient expression, and on A. rhizogenes R1000 for its hairy root induction frequency. For AGL1, Silwet S-408 produced higher level of expression than Silwet L-77, were 2.3- and 1.3-fold, respectively. For GV3101, Silwet L-77 was still higher than Silwet S-408, were 1.5- and 1.4-fold, respectively. However, GV3101 produced higher levels of expression than AGL1. The area of GUS expression spots of AGL1, LBA4404, and GV3101 strains was 53.43%, 41.06%, and 30.51%, respectively.Conclusion:A. tumefaciens AGl1 strain was the most effective to be transformed in to A. annua than GV3101 and LBA4404 strain. Surfactant Silwet S-408 produced the highest efficiency of transformation.

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“…The microbes used in the metabolic engineering of artemisinin include Escherichia coli (Martin et al, 2003), Saccharomyces cerevisiae (Ro et al, 2006), Aspergillus nidulans (Lubertozzi and Keasling, 2008) and others. S. cerevisiae, an eukaryotic microbe, is an attractive host for the production of artemisinin precursors because it can produce farnesyl pyrophosphate for its sterol biosynthesis.…”

Section: Introductionmentioning

confidence: 99%

Improvement of Amorpha-4,11-diene Production by a Yeast-Conform Variant of Vitreoscilla Hemoglobin

Shen¹,

Zhi²,

Shi³

et al. 2012

Z. Naturforsch. C

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Amorpha-4,11-diene is the precursor of the antimalarial compound artemisinin. The effect of Vitreoscilla hemoglobin (VHb) and its yeast-conform variant (VHbm) on amorpha-4,11diene production in engineered Saccharomyces cerevisiae was investigated. First, the VHb gene was mutated to the yeast-conform variant VHbm based on step-by-step extension of a short region of the gene through a series of polymerase chain reactions (PCR). The artifi cial VHbm gene contained codons preferred by the yeast translation machinery. Two yeast expression vectors containing VHb or VHbm gene were constructed and introduced into the amorpha-4,11-diene-producing strain S. cerevisiae WK1 to form WK1[VHb] and WK1[VHbm], respectively. Western blot and CO-difference spectrum absorbance assay showed that VHb and VHbm were successfully expressed. In shake fl asks, VHbm expression conferred higher cell growth than VHb expression. GC-MS results indicated the amorpha-4,11-diene production in WK1[VHbm] and WK1[VHb] was 3-and 2-fold higher than that in WK1, respectively. This suggests that VHb might improve the amorpha-4,11-diene production in engineered S. cerevisiae.

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Expression of a synthetic Artemesia annua amorphadiene synthase in Aspergillus nidulans yields altered product distribution

Cited by 19 publications

References 31 publications

Functional Expression of an Orchid Fragrance Gene in Lactococcus lactis

Functional Expression of an Orchid Fragrance Gene in Lactococcus lactis

Optimization of genetic transformation of Artemisia annua L. Using Agrobacterium for Artemisinin production

Improvement of Amorpha-4,11-diene Production by a Yeast-Conform Variant of Vitreoscilla Hemoglobin

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