Dataset Information

Isolation of thermotolerant yeast Pichia kudriavzevii from nuruk.

ABSTRACT: Thermotolerant yeast strains were isolated from nuruk, a traditional Korean fermentation starter in which variety of microorganisms are present. Among the isolates, the MBY1358 identified as yeast Pichia kudriavzevii showed significantly higher growth rate (0.59 ± 0.00 1/h) at 44 °C than other strains. Maximum ethanol concentration of 8.35 ± 0.03 g/L was obtained from 20 g/L glucose with yield of 0.44 ± 0.01 g/g at 44 °C, which is 1.14 times ethanol production of the control strain of P. kudriavzevii. The MBY1358, which was significantly more thermotolerant than the control strain and fermented 200 g/L glucose to 107.33 ± 5.03 g/L ethanol at 44 °C, was deposited to Korean Collection for Type Cultures (KCTC) under the accession number 27654.

SUBMITTER: Choi DH

PROVIDER: S-EPMC6049779 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Isolation of thermotolerant yeast Pichia kudriavzevii from nuruk.

Choi Da-Hye DH Park Eun-Hee EH Kim Myoung-Dong MD

Food science and biotechnology 20170818 5

Thermotolerant yeast strains were isolated from nuruk, a traditional Korean fermentation starter in which variety of microorganisms are present. Among the isolates, the MBY1358 identified as yeast Pichia kudriavzevii showed significantly higher growth rate (0.59 ± 0.00 1/h) at 44 °C than other strains. Maximum ethanol concentration of 8.35 ± 0.03 g/L was obtained from 20 g/L glucose with yield of 0.44 ± 0.01 g/g at 44 °C, which is 1.14 times ethanol production of the control strain ...[more]

PMID: 30263670

Similar Datasets

Project description:Itaconic acid (IA), or 2-methylenesuccinic acid, has a broad spectrum of applications in the biopolymer industry owing to the presence of one vinyl bond and two acid groups in the structure. Its polymerization can follow a similar mechanism as acrylic acid but additional functionality can be incorporated into the extra beta acid group. Currently, the bio-based production of IA in industry relies on the fermentation of the filamentous fungus Aspergillus terreus. However, the difficulties associated with the fermentation undertaken by filamentous fungi together with the pathogenic potential of A. terreus pose a serious challenge for industrial-scale production. In recent years, there has been increasing interest in developing alternative production hosts for fermentation processes that are more homogenous in the production of organic acids. Pichia kudriavzevii is a non-conventional yeast with high acid tolerance to organic acids at low pH, which is a highly desirable trait by easing downstream processing. We introduced cis-aconitic acid decarboxylase gene (cad) from A. terreus (designated At_cad) into this yeast and established the initial titer of IA at 135 ± 5 mg/L. Subsequent overexpression of a native mitochondrial tricarboxylate transporter (herein designated Pk_mttA) presumably delivered cis-aconitate efficiently to the cytosol and doubled the IA production. By introducing the newly invented CRISPR-Cas9 system into P. kudriavzevii, we successfully knocked out both copies of the gene encoding isocitrate dehydrogenase (ICD), aiming to increase the availability of cis-aconitate. The resulting P. kudriavzevii strain, devoid of ICD and overexpressing Pk_mttA and At_cad on its genome produced IA at 505 ± 17.7 mg/L in shake flasks, and 1232 ± 64 mg/L in fed-batch fermentation. Because the usage of an acid-tolerant species does not require pH adjustment during fermentation, this work demonstrates the great potential of engineering P. kudriavzevii as an industrial chassis for the production of organic acid.

Project description:The unconventional yeast Pichia kudriavzevii is renowned for its ability to survive at low pH and has been exploited for the industrial production of various organic acids, especially succinic acid (SA). However, P. kudriavzevii can also utilize the di- and tricarboxylate intermediates of the Krebs cycle as the sole carbon sources for cell growth, which may adversely affect the extracellular accumulation of SA. Because the carboxylic acid transport machinery of P. kudriavzevii remains poorly understood, here, we focused on studying its SA transportation process from the perspective of mining and characterization of dicarboxylate transporters in a newly isolated acid-tolerant P. kudriavzevii strain CY902. Through genome sequencing and transcriptome analysis, two JEN family carboxylate transporters (PkJEN2-1 and PkJEN2-2) were found to be involved in SA transport. Substrate specificity analysis revealed that both PkJEN proteins are active dicarboxylate transporters, that can effectively import succinate, fumarate and L-malate into the cell. In addition, PkJEN2-1 can transport α-ketoglutarate, while PkJEN2-2 cannot. Since PkJEN2-1 shows higher transcript abundance than PkJEN2-2, its role in dicarboxylate transport is more important than PkJEN2-2. In addition, PKJEN2-2 is also responsible for the uptake of citrate. To our best knowledge, this is the first study to show that a JEN2 subfamily transporter is involved in tricarboxylate transport in yeast. A combination of model-based structure analysis and rational mutagenesis further proved that amino acid residues 392-403 of the tenth transmembrane span (TMS-X) of PkJEN2-2 play an important role in determining the specificity of the tricarboxylate substrate. Moreover, these two PkJEN transporters only exhibited inward transport activity for SA, and simultaneous inactivation of both PkJEN transporters reduced the SA influx, resulting in enhanced extracellular accumulation of SA in the late stage of fermentation. This work provides useful information on the mechanism of di-/tricarboxylic acid utilization in P. kudriavzevii, which will help improve the organic acid production performance of this microbial chassis.

Dataset Information

Isolation of thermotolerant yeast Pichia kudriavzevii from nuruk.

Publications

Isolation of thermotolerant yeast <i>Pichia kudriavzevii</i> from <i>nuruk</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets