Impact of culture conditions on β-carotene encapsulation using Yarrowia lipolytica cells

2017 ◽  
Author(s):  
Tran Hai Dang ◽  
Ho Thi Thu Minh ◽  
Tran Nguyen Van Nhi ◽  
Ta Thi Minh Ngoc
Keyword(s):  
Yarrowia Lipolytica ◽  
Culture Conditions ◽  
Β Carotene

scholarly journals Aeration and Stirring in Yarrowia lipolytica Lipase Biosynthesis during Batch Cultures with Waste Fish Oil as a Carbon Source

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 88
Author(s):  
Paulina Snopek ◽  
Dorota Nowak ◽  
Bartłomiej Zieniuk ◽  
Agata Fabiszewska
Keyword(s):  
Carbon Source ◽  
Fish Oil ◽  
Yarrowia Lipolytica ◽  
Lipolytic Activity ◽  
Culture Conditions ◽  
Agitation Speed ◽  
Oxygen Deficit ◽  
Shear Stresses ◽  
Process Selection ◽  
Batch Cultures

Yarrowia lipolytica is one of the most studied non-conventional forms of yeast, exhibiting a high secretory capacity and producing many industrially important and valuable metabolites. The yeast conceals a great biotechnological potential to synthesize organic acids, sweeteners, microbial oil, or fragrances. The vast majority of bioprocesses are carried out in bioreactors, where suitable culture conditions are provided. In the current study, the effect of agitation speed (200–600 rpm) and air flow rate (0.0375–2.0 dm3/(dm3 × min)) on the biomass yield and lipase activity of Y. lipolytica KKP 379 is analyzed in a growth medium containing waste fish oil. The increase of aeration intensity limited the period of oxygen deficit in the medium. Simultaneously, an increase in lipolytic activity was observed from 2.09 U/cm3 to 14.21 U/cm3; however, an excessive agitation speed likely caused oxidative or shear stresses, and a reduction in lipolytic activity was observed. Moreover, it is confirmed that the synthesis of lipases is related to oxygen consumption, pH, and the yeast growth phase, and appropriate process selection may provide two advantages, namely, the maximum use of the waste carbon source and the production of lipolytic enzymes that are valuable in many industries.

scholarly journals Metabolic Engineering of Non-carotenoid-Producing Yeast Yarrowia lipolytica for the Biosynthesis of Zeaxanthin

2021 ◽  
Vol 12 ◽  
Author(s):  
Yuxiao Xie ◽  
Shulin Chen ◽  
Xiaochao Xiong
Keyword(s):  
Yarrowia Lipolytica ◽  
Fold Increase ◽  
Genetically Engineered ◽  
Cell Factory ◽  
Yeast Nitrogen Base ◽  
Nitrogen Base ◽  
Yeast Extract Peptone Dextrose ◽  
Dry Cell Weight ◽  
A Cell ◽  
Β Carotene

Zeaxanthin is vital to human health; thus, its production has received much attention, and it is also an essential precursor for the biosynthesis of other critical carotenoids such as astaxanthin and crocetin. Yarrowia lipolytica is one of the most intensively studied non-conventional yeasts and has been genetically engineered as a cell factory to produce carotenoids such as lycopene and β-carotene. However, zeaxanthin production by Y. lipolytica has not been well investigated. To fill this gap, β-carotene biosynthesis pathway has been first constructed in this study by the expression of genes, including crtE, crtB, crtI, and carRP. Three crtZ genes encoding β-carotene hydroxylase from different organisms were individually introduced into β-carotene-producing Y. lipolytica to evaluate their performance for producing zeaxanthin. The expression of crtZ from the bacterium Pantoea ananatis (formerly Erwinia uredovora, Eu-crtZ) resulted in the highest zeaxanthin titer and content on the basis of dry cell weight (DCW). After verifying the function of Eu-crtZ for producing zeaxanthin, the high-copy-number integration into the ribosomal DNA of Y. lipolytica led to a 4.02-fold increase in the titer of zeaxanthin and a 721% increase in the content of zeaxanthin. The highest zeaxanthin titer achieved 21.98 ± 1.80 mg/L by the strain grown on a yeast extract peptone dextrose (YPD)–rich medium. In contrast, the highest content of DCW reached 3.20 ± 0.11 mg/g using a synthetic yeast nitrogen base (YNB) medium to culture the cells. Over 18.0 g/L of citric acid was detected in the supernatant of the YPD medium at the end of cultivation. Furthermore, the zeaxanthin-producing strains still accumulated a large amount of lycopene and β-carotene. The results demonstrated the potential of a cell factory for zeaxanthin biosynthesis and opened up an avenue to engineer this host for the overproduction of carotenoids.

Metabolic engineering of β-carotene biosynthesis in Yarrowia lipolytica

2020 ◽  
Vol 42 (6) ◽  
pp. 945-956 ◽  
Author(s):  
Xin-Kai Zhang ◽  
Dan-Ni Wang ◽  
Jun Chen ◽  
Zhi-Jie Liu ◽  
Liu-Jing Wei ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Yarrowia Lipolytica ◽  
Β Carotene

scholarly journals Enhancement of Astaxanthin Biosynthesis in Oleaginous Yeast Yarrowia lipolytica via Microalgal Pathway

2019 ◽  
Vol 7 (10) ◽  
pp. 472 ◽  
Author(s):  
Larissa Ribeiro Ramos Tramontin ◽  
Kanchana Rueksomtawin Kildegaard ◽  
Suresh Sudarsan ◽  
Irina Borodina
Keyword(s):  
Yarrowia Lipolytica ◽  
Oleaginous Yeast ◽  
Small Scale ◽  
Cell Factory ◽  
Standard Equipment ◽  
Cell Factories ◽  
Dry Cell Weight ◽  
Astaxanthin Production ◽  
Yeast Yarrowia Lipolytica ◽  
Β Carotene

Astaxanthin is a high-value red pigment and antioxidant used by pharmaceutical, cosmetics, and food industries. The astaxanthin produced chemically is costly and is not approved for human consumption due to the presence of by-products. The astaxanthin production by natural microalgae requires large open areas and specialized equipment, the process takes a long time, and results in low titers. Recombinant microbial cell factories can be engineered to produce astaxanthin by fermentation in standard equipment. In this work, an oleaginous yeast Yarrowia lipolytica was engineered to produce astaxanthin at high titers in submerged fermentation. First, a platform strain was created with an optimised pathway towards β-carotene. The platform strain produced 331 ± 66 mg/L of β-carotene in small-scale cultivation, with the cellular content of 2.25% of dry cell weight. Next, the genes encoding β-ketolase and β-hydroxylase of bacterial (Paracoccus sp. and Pantoea ananatis) and algal (Haematococcus pluvialis) origins were introduced into the platform strain in different copy numbers. The resulting strains were screened for astaxanthin production, and the best strain, containing algal β-ketolase and β-hydroxylase, resulted in astaxanthin titer of 44 ± 1 mg/L. The same strain was cultivated in controlled bioreactors, and a titer of 285 ± 19 mg/L of astaxanthin was obtained after seven days of fermentation on complex medium with glucose. Our study shows the potential of Y. lipolytica as the cell factory for astaxanthin production.

Strain and Culture Conditions Improvement for β-Carotene Production With Mucor

2005 ◽  
pp. 239-256 ◽  
Author(s):  
Enrique A. ◽  
Tamás Papp ◽  
Jesper Breum ◽  
José Arnau ◽  
Arturo P.
Keyword(s):  
Culture Conditions ◽  
Β Carotene

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

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.

Accumulation of a Novel Red Pigment in Cell Suspension Cultures of Floral Meristem Tissues from Carthamus tinctorius L.

1988 ◽  
Vol 43 (11-12) ◽  
pp. 862-870 ◽  
Author(s):  
Koshi Saito ◽  
Etuko Daimon ◽  
Kazuhito Kusaka ◽  
Sachio Wakayama ◽  
Yoshihiro Sekino
Keyword(s):  
Cell Suspension ◽  
Culture Media ◽  
Cell Suspension Cultures ◽  
Carthamus Tinctorius ◽  
Culture Conditions ◽  
Floral Meristem ◽  
Red Pigment ◽  
A Cell ◽  
Micro Elements ◽  
Β Carotene

Abstract A cell strain originally derived from floral meristem tissues of Carthamus tinctorius (dyer’s saffron) produced substantial amounts of a novel red pigment under controlled culture conditions. The pigment isolated f r om alcoholic extracts of C. tinctorius cultures was compared with authentic carthamin, anthocyanins, betain, and carotenoids. It differed markedly from carthamin and showed none of the characteristic properties of the glycoside or chloride forms of authentic delphinidin, cyanidin, and pelargonidin. Analytic data indicated that this pigment also differs from betanin and from α- and β-carotene. The name “Kurenamin” was tentatively given to this newly isolated red pigment. Effect of the culture media, micro-elements, macro-elements, and putative substrates on the kurenamin production were investigated during cell suspension culture.

scholarly journals High-Level Production of a Thermostable Mutant of Yarrowia lipolytica Lipase 2 in Pichia pastoris

2019 ◽  
Vol 21 (1) ◽  
pp. 279
Author(s):  
Qinghua Zhou ◽  
Zhixin Su ◽  
Liangcheng Jiao ◽  
Yao Wang ◽  
Kaixin Yang ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Yarrowia Lipolytica ◽  
Large Scale ◽  
Gene Dosage ◽  
Life Time ◽  
Industrial Applications ◽  
Culture Conditions ◽  
Scale Production ◽  
Large Scale Production ◽  
High Level

As a promising biocatalyst, Yarrowia lipolytica lipase 2 (YlLip2) is limited in its industrial applications due to its low thermostability. In this study, a thermostable YlLip2 mutant was overexpressed in Pichia pastoris and its half-life time was over 30 min at 80 °C. To obtain a higher protein secretion level, the gene dosage of the mutated lip2 gene was optimized and the lipase activity was improved by about 89%. Then, the YlLip2 activity of the obtained strain further increased from 482 to 1465 U/mL via optimizing the shaking flask culture conditions. Subsequently, Hac1p and Vitreoscilla hemoglobin (VHb) were coexpressed with the YlLip2 mutant to reduce the endoplasmic reticulum stress and enhance the oxygen uptake efficiency in the recombinant strains, respectively. Furthermore, high-density fermentations were performed in a 3 L bioreactor and the production of the YlLip2 mutant reached 9080 U/mL. The results demonstrated that the expression level of the thermostable YlLip2 mutant was predominantly enhanced via the combination of these strategies in P. pastoris, which forms a consolidated basis for its large-scale production and future industrial applications.

Reexamination of the Carotenoid Pigments of the Unicellular Blue-green Alga Agmenellum quadruplicatum

1977 ◽  
Vol 34 (5) ◽  
pp. 659-668 ◽  
Author(s):  
N. J. Antia ◽  
J. Y. Cheng
Keyword(s):  
Culture Conditions ◽  
Carotenoid Pigments ◽  
Chlorophyll B ◽  
Blue Green Alga ◽  
Typical Member ◽  
Agmenellum Quadruplicatum ◽  
Carotenoid Composition ◽  
Previous Claim ◽  
Consecutive Loss ◽  
Β Carotene

A previous claim for the occurrence of antheraxanthin and the absence of typical cyanophycean xanthophylls in Agmenellum quadruplicatum has been disproven. This phycocyanin-producing prokaryote showed chlorophyll a, β-carotene, zeaxanthin, one myxoxanthophyll-like, and one myxol-like carotenoid as the major pigments. Traces of keto and other carotenoids were observed, one of which appears to be phoenicopterone or a cis-echinenone. No antheraxanthin and chlorophyll b or c were detected under all culture conditions tested. Algal chlorosis from culture aging was caused by the consecutive loss of phycocyanin and chlorophyll, but the carotenoid composition was much less affected and there was no gain of secondary or keto carotenoids. We concluded that this alga is a typical member of the Cyanophyceae in terms of pigment chemotaxonomy.

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