Protoplast fusion between Blakeslea trispora 14,271 (+) and 14,272 (−) enhanced the yield of lycopene and β-carotene

ABSTRACT We cloned the carB and carRA genes involved in β-carotene biosynthesis from overproducing and wild-type strains of Blakeslea trispora. The carB gene has a length of 1,955 bp, including two introns of 141 and 68 bp, and encodes a protein of 66.4 kDa with phytoene dehydrogenase activity. The carRA gene contains 1,894 bp, with a single intron of 70 bp, and encodes a protein of 69.6 kDa with separate domains for lycopene cyclase and phytoene synthase. The estimated transcript sizes for carB and carRA were 1.8 and 1.9 kb, respectively. CarB from the β-carotene-overproducing strain B. trispora F-744 had an S528R mutation and a TAG instead of a TAA stop codon. The overproducing strain also had a P143S mutation in CarRA. Both B. trispora genes could complement mutations in orthologous genes in Mucor circinelloides and could be used to construct transformed strains of M. circinelloides that produced higher levels of β-carotene than did the nontransformed parent. The results show that these genes are conserved across the zygomycetes and that the B. trispora carB and carRA genes are functional and potentially useable to increase carotenoid production.

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Enhanced synthesis of trisporic acid and β-carotene production in Blakeslea trispora by addition of a non-ionic surfactant, Span 20

Journal of Fermentation and Bioengineering ◽

10.1016/s0922-338x(97)89253-7 ◽

1997 ◽

Vol 84 (4) ◽

pp. 330-332 ◽

Cited By ~ 27

Author(s):

Seon-Won Kim ◽

Weon-Taek Seo ◽

Young-Hoon Park

Keyword(s):

Blakeslea Trispora ◽

Ionic Surfactant ◽

Β Carotene

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Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica

Frontiers in Microbiology ◽

10.3389/fmicb.2021.627150 ◽

2021 ◽

Vol 12 ◽

Author(s):

Liang Liu ◽

Yu Ling Qu ◽

Gui Ru Dong ◽

Jing Wang ◽

Ching Yuan Hu ◽

...

Keyword(s):

Yarrowia Lipolytica ◽

Large Scale ◽

Blakeslea Trispora ◽

Scale Production ◽

Total Production ◽

Metabolic Balance ◽

Carotene Content ◽

Large Scale Production ◽

Dry Cell Weight ◽

Β Carotene

β-carotene is a precursor of vitamin A and has multiple physiological functions. Producing β-carotene by microbial fermentation has attracted much attention to consumers’ preference for natural products. This study focused on improving β-carotene production by constructing codon-adapted genes and minimizing intermediate accumulation. The codon-adapted CarRA and CarB genes from the industrial strain of Blakeslea trispora were integrated into the genome of the Yarrowia lipolytica to construct YL-C0, the baseline strain for producing β-carotene. Thereafter, the β-carotene biosynthetic pathway’s metabolic balance was accurately regulated to reduce the intermediates’ accumulation. Notably, the β-carotene content increased by 21 times to reach 12.5 dry cell weight (DCW) mg/g when minimizing HMG-CoA and FPP accumulation. Further, we improved the expression levels of the CarRA and CarB genes to minimize the accumulation of phytoene and lycopene. Total production of β-carotene of 1.7 g/L and 21.6 mg/g DCW was achieved. These results reveal that the rate-limiting enzymes CarRA and CarB of B. trispora exhibited higher catalytic activity than the same enzymes from other microorganisms. Promoting metabolic balance by minimizing the accumulation of intermediates is a very effective strategy for increasing β-carotene. The β-carotene-producing strain constructed in this study has established the foundation for its potential use in industrial production. These successful engineering strategies also provide a foundation for large-scale production of other terpenoids.

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Medium optimization and downstream process design for the augmented yield of β-Carotene using fungi Blakeslea trispora

Pigment & Resin Technology ◽

10.1108/prt-05-2021-0055 ◽

2021 ◽

Vol ahead-of-print (ahead-of-print) ◽

Author(s):

Umesh Luthra ◽

Prabhakar Babu ◽

Remya R.R. ◽

Angeline Julius ◽

Yogesh Patel ◽

...

Keyword(s):

Process Design ◽

Medium Optimization ◽

Corn Starch ◽

Uv Spectroscopy ◽

Maximum Activity ◽

Blakeslea Trispora ◽

Content Type ◽

Natural Oils ◽

Lower Temperature ◽

Β Carotene

Purpose β-Carotene is the most appropriate and significant precursor of vitamin A. Synthetic carotene supplements have been known to pose a threat to human health, making natural sources such as the indefensible choice for the production and extraction of carotene. Design/methodology/approach This study considers Blakeslea trispora, a filamentous fungus, as a source of production of carotenoids by fermentation and wet and dry mycelium were used to analyse and obtain better extraction results. Findings In this study, natural oils such as soy oil and cottonseed oil were incorporated into fermentation media to increase the production of carotene. For the optimization process, Plackett–Burman and one-factor-at-a-time (OVAT) models were identified as being of great value. Originality/value OVAT was carried out for corn starch because it plays a major role in the production of carotene and the corn starch at 30 g/L concentration has shown the maximum activity of 3.48 mg/gm. After optimizing process variables, submerged fermentation was eventually carried out under highly controlled media conditions. The resulting product was quantified using UV spectroscopy and extraction of carotene has been observed in the presence of various solvents. Among a range of solvents used, the methylene Di chloride produced-carotene at 86% recovery at a significantly lower temperature of 35°C.

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Optimizing β-carotene production by Blakeslea trispora using bug damaged wheat

Pigment & Resin Technology ◽

10.1108/prt-05-2017-0055 ◽

2018 ◽

Vol 47 (3) ◽

pp. 189-195

Author(s):

Serap Durakli Velioglu ◽

Goksel Tirpanci Sivri

Keyword(s):

Response Surface Methodology ◽

Industrial Waste ◽

Design Methodology ◽

Model Fit ◽

Quadratic Model ◽

Blakeslea Trispora ◽

Natural Food ◽

Control Medium ◽

Content Type ◽

Β Carotene

Purpose β-carotene, found naturally in many foods, has been widely used as a colourant in various products. The aim of this study is to optimize the production of β-carotene by Blakeslea trispora using bug-damaged wheat as the main nitrogen and carbon source. Design/methodology/approach Response surface methodology (RSM) has been used to determine the optimum concentrations of media components for the maximum β-carotene production. β-carotene concentration was determined quantitively using a spectrophotometric method. Findings A full quadratic model fit the data best for β-carotene production with a high R2 value (0.999). Using optimized media increased the production of β-carotene by B. trispora DSM-2387 and B. trispora DSM-2388 to a higher level (250.407 mg/l) than the control medium (72.195 mg/l). Originality/value This study showed that bug-damaged wheat, an agro-industrial waste having low economical value, may be used as a valuable substrate for microbial production of β-carotene, a natural food colourant possessing high economical value.

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