Carbon source requirements for mating and mating‐type switching in the methylotrophic yeasts Ogataea (Hansenula) polymorpha and Komagataella phaffii (Pichia pastoris)

Yeast ◽  
2020 ◽  
Vol 37 (2) ◽  
pp. 237-245 ◽  
Author(s):  
Dahao Feng ◽  
Anton Stoyanov ◽  
Juliana C. Olliff ◽  
Kenneth H. Wolfe ◽  
Kantcho Lahtchev ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Carbon Source ◽  
Mating Type ◽  
Hansenula Polymorpha ◽  
Methylotrophic Yeasts ◽  
Komagataella Phaffii ◽  
Mating Type Switching

scholarly journals Microbe Profile: Komagataella phaffii: a methanol devouring biotech yeast formerly known as Pichia pastoris

Microbiology ◽  
2020 ◽  
Vol 166 (7) ◽  
pp. 614-616 ◽  
Author(s):  
Lina Heistinger ◽  
Brigitte Gasser ◽  
Diethard Mattanovich
Keyword(s):  
Energy Source ◽  
Pichia Pastoris ◽  
Secondary Metabolism ◽  
Mating Type ◽  
Heterologous Proteins ◽  
Methylotrophic Yeasts ◽  
Komagataella Phaffii ◽  
Primary And Secondary Metabolism ◽  
Diploid Cells

Methylotrophic yeasts of the genus Komagataella are abundantly found in tree exudates. Their ability to utilize methanol as carbon and energy source relies on an assimilation pathway localized in largely expanded peroxisomes, and a cytosolic methanol dissimilation pathway. Other substrates like glucose or glycerol are readily utilized as well. Komagataella yeasts usually grow as haploid cells and are secondary homothallic as they can switch mating type. Upon mating diploid cells sporulate readily, forming asci with four haploid spores. Their ability to secrete high amounts of heterologous proteins made them interesting for biotechnology, which expands today also to other products of primary and secondary metabolism.

Isolation of alcohol oxidase and two other methanol regulatable genes from the yeast Pichia pastoris

1985 ◽  
Vol 5 (5) ◽  
pp. 1111-1121
Author(s):  
S B Ellis ◽  
P F Brust ◽  
P J Koutz ◽  
A F Waters ◽  
M M Harpold ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Alcohol Oxidase ◽  
Methylotrophic Yeasts ◽  
Sequence Analyses ◽  
Yeast Pichia Pastoris ◽  
Oxidation Of Methanol ◽  
Regulated Expression

The oxidation of methanol follows a well-defined pathway and is similar for several methylotrophic yeasts. The use of methanol as the sole carbon source for the growth of Pichia pastoris stimulates the expression of a family of genes. Three methanol-responsive genes have been isolated; cDNA copies have been made from mRNAs of these genes, and the protein products from in vitro translations have been examined. The identification of alcohol oxidase as one of the cloned, methanol-regulated genes has been made by enzymatic, immunological, and sequence analyses. Methanol-regulated expression of each of these three isolated genes can be demonstrated to occur at the level of transcription. Finally, DNA subfragments of two of the methanol-responsive genomic clones from P. pastoris have been isolated and tentatively identified as containing the control regions involved in methanol regulation.

scholarly journals Establishment of a co-culture system using Escherichia coli and Pichia pastoris (Komagataella phaffii) for valuable alkaloid production

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Miya Urui ◽  
Yasuyuki Yamada ◽  
Yoshito Ikeda ◽  
Akira Nakagawa ◽  
Fumihiko Sato ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pichia Pastoris ◽  
Carbon Source ◽  
Culture System ◽  
De Novo ◽  
Plant Metabolites ◽  
E Coli ◽  
Komagataella Phaffii ◽  
Pharmaceutical Industries ◽  
Valuable Compounds

Abstract Background Plants produce a variety of specialized metabolites, many of which are used in pharmaceutical industries as raw materials. However, certain metabolites may be produced at markedly low concentrations in plants. This problem has been overcome through metabolic engineering in recent years, and the production of valuable plant compounds using microorganisms such as Escherichia coli or yeast cells has been realized. However, the development of complicated pathways in a single cell remains challenging. Additionally, microbial cells may experience toxicity from the bioactive compounds produced or negative feedback effects exerted on their biosynthetic enzymes. Thus, co-culture systems, such as those of E. coli–E. coli and E. coli-Saccharomyces cerevisiae, have been developed, and increased production of certain compounds has been achieved. Recently, a co-culture system of Pichia pastoris (Komagataella phaffii) has gained considerable attention due to its potential utility in increased production of valuable compounds. However, its co-culture with other organisms such as E. coli, which produce important intermediates at high concentrations, has not been reported. Results Here, we present a novel co-culture platform for E. coli and P. pastoris. Upstream E. coli cells produced reticuline from a simple carbon source, and the downstream P. pastoris cells produced stylopine from reticuline. We investigated the effect of four media commonly used for growth and production of P. pastoris, and found that buffered methanol-complex medium (BMMY) was suitable for P. pastoris cells. Reticuline-producing E. coli cells also showed better growth and reticuline production in BMMY medium than that in LB medium. De novo production of the final product, stylopine from a simple carbon source, glycerol, was successful upon co-culture of both strains in BMMY medium. Further analysis of the initial inoculation ratio showed that a higher ratio of E. coli cells compared to P. pastoris cells led to higher production of stylopine. Conclusions This is the first report of co-culture system established with engineered E. coli and P. pastoris for the de novo production of valuable compounds. The co-culture system established herein would be useful for increased production of heterologous biosynthesis of complex specialized plant metabolites.

scholarly journals Isolation of alcohol oxidase and two other methanol regulatable genes from the yeast Pichia pastoris.

1985 ◽  
Vol 5 (5) ◽  
pp. 1111-1121 ◽  
Author(s):  
S B Ellis ◽  
P F Brust ◽  
P J Koutz ◽  
A F Waters ◽  
M M Harpold ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Alcohol Oxidase ◽  
Methylotrophic Yeasts ◽  
Sequence Analyses ◽  
Yeast Pichia Pastoris ◽  
Oxidation Of Methanol ◽  
Regulated Expression

The oxidation of methanol follows a well-defined pathway and is similar for several methylotrophic yeasts. The use of methanol as the sole carbon source for the growth of Pichia pastoris stimulates the expression of a family of genes. Three methanol-responsive genes have been isolated; cDNA copies have been made from mRNAs of these genes, and the protein products from in vitro translations have been examined. The identification of alcohol oxidase as one of the cloned, methanol-regulated genes has been made by enzymatic, immunological, and sequence analyses. Methanol-regulated expression of each of these three isolated genes can be demonstrated to occur at the level of transcription. Finally, DNA subfragments of two of the methanol-responsive genomic clones from P. pastoris have been isolated and tentatively identified as containing the control regions involved in methanol regulation.

scholarly journals Centromeres of the yeast Komagataella phaffii (Pichia pastoris) have a simple inverted-repeat structure

2016 ◽  
Author(s):  
Aisling Y. Coughlan ◽  
Sara J. Hanson ◽  
Kevin P. Byrne ◽  
Kenneth H. Wolfe
Keyword(s):  
Candida Albicans ◽  
Pichia Pastoris ◽  
Inverted Repeat ◽  
Methylotrophic Yeast ◽  
H3k9 Methylation ◽  
Komagataella Phaffii ◽  
Repeat Structure ◽  
Different Types ◽  
Mating Type Switching ◽  
The Relationship

AbstractCentromere organization has evolved dramatically in one clade of fungi, the Saccharomycotina. These yeasts have lost the ability to make normal eukaryotic heterochromatin with histone H3K9 methylation, which is a major component of pericentromeric regions in other eukaryotes. Following this loss, several different types of centromere emerged, including two types of sequence-defined ("point") centromeres, and the epigenetically-defined "small regional" centromeres of Candida albicans. Here we report that centromeres of the methylotrophic yeast Komagataella phaffii (formerly called Pichia pastoris) are structurally-defined. Each of its four centromeres consists of a 2-kb inverted repeat (IR) flanking a 1-kb central core (mid) region. The four centromeres are unrelated in sequence. CenH3 (Cse4) binds strongly to the cores, with a decreasing gradient along the IRs. This mode of organization resembles Schizosaccharomyces pombe centromeres but is much more compact and lacks the extensive flanking heterochromatic otr repeats. Different isolates of K. phaffii show polymorphism for the orientation of the mid regions, due to recombination in the IRs. CEN4 is located within a 138-kb region that changes orientation during mating-type switching, but switching does not induce recombination of centromeric IRs. The existing genetic toolbox for K. phaffii should facilitate analysis of the relationship between the IRs and the establishment and maintenance of centromeres in this species.

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