β-Galactosidase over-production by a mig1 mutant of Kluyveromyces marxianus KM for efficient hydrolysis of lactose

Zhou, Hai‐Xiang; Xu, Jin-Li; Chi, Zhe; Liu, Guanglei

doi:10.1016/j.bej.2013.04.010

Cited by 21 publications

(8 citation statements)

References 21 publications

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“…Therefore, KmHXK1 and KmSNF1 were the key genes involved in the glucose repression pathway of K. marxianus . Km MIG1 was involved in the glucose repression on nonglucose sugars like galactose in K. marxianus , which is consistent with previous study [35]. Nonetheless, strain with Km MIG1 disruption did not release the utilization of xylose from glucose repression.…”

Section: Resultssupporting

confidence: 92%

Release of glucose repression on xylose utilization in Kluyveromyces marxianus to enhance glucose-xylose co-utilization and xylitol production from corncob hydrolysate

et al. 2019

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BackgroundLignocellulosic biomass is one of the most abundant materials for biochemicals production. However, efficient co-utilization of glucose and xylose from the lignocellulosic biomass is a challenge due to the glucose repression in microorganisms. Kluyveromyces marxianus is a thermotolerant and efficient xylose-utilizing yeast. To realize the glucose–xylose co-utilization, analyzing the glucose repression of xylose utilization in K. marxianus is necessary. In addition, a glucose–xylose co-utilization platform strain will facilitate the construction of lignocellulosic biomass-utilizing strains.ResultsThrough gene disruption, hexokinase 1 (KmHXK1) and sucrose non-fermenting 1 (KmSNF1) were proved to be involved in the glucose repression of xylose utilization while disruption of the downstream genes of cyclic AMP-protein kinase A (cAMP-PKA) signaling pathway or sucrose non-fermenting 3 (SNF3) glucose-sensing pathway did not alleviate the repression. Furthermore, disruption of the gene of multicopy inhibitor of GAL gene expression (KmMIG1) alleviated the glucose repression on some nonglucose sugars (galactose, sucrose, and raffinose) but still kept glucose repression of xylose utilization. Real-time PCR analysis of the xylose utilization related genes transcription confirmed these results, and besides, revealed that xylitol dehydrogenase gene (KmXYL2) was the critical gene for xylose utilization and stringently regulated by glucose repression. Many other genes of candidate targets interacting with SNF1 were also evaluated by disruption, but none proved to be the key regulator in the pathway of the glucose repression on xylose utilization. Therefore, there may exist other signaling pathway(s) for glucose repression on xylose consumption. Based on these results, a thermotolerant xylose–glucose co-consumption platform strain of K. marxianus was constructed. Then, exogenous xylose reductase and xylose-specific transporter genes were overexpressed in the platform strain to obtain YHY013. The YHY013 could efficiently co-utilized the glucose and xylose from corncob hydrolysate or xylose mother liquor for xylitol production (> 100 g/L) even with inexpensive organic nitrogen sources.ConclusionsThe analysis of the glucose repression in K. marxianus laid the foundation for construction of the glucose–xylose co-utilizing platform strain. The efficient xylitol production strain further verified the potential of the platform strain in exploitation of lignocellulosic biomass.Electronic supplementary materialThe online version of this article (10.1186/s12934-019-1068-2) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 92%

Release of glucose repression on xylose utilization in Kluyveromyces marxianus to enhance glucose-xylose co-utilization and xylitol production from corncob hydrolysate

et al. 2019

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“…The medium for pullulan production consisted of 138.0 g/l inulin, 3.0 g/l yeast extract, 5.0 g/l K 2 HPO 4 , 0.2 g/l MgSO 4 ·7H 2 O, 1.0 g/l NaCl, and 0.6 g/l (NH 4 ) 2 SO 4 (Duan et al 2008). K. maximum KM which is a high inulinase producer was grown in the inulinase production medium (Zhou et al 2013). The Escherichia coli strain DH5α [F − endA1 hsdR17 (rK_/mK + ) supE44 thi − λ − recA1gyr96DlacU169 (j80lacZDM15)] was used in this study and was grown in Luria-Bertani broth (LB).…”

Section: Strains and Cultivation Mediamentioning

confidence: 99%

Genetic Modification of the Marine-Isolated Yeast Aureobasidium melanogenum P16 for Efficient Pullulan Production from Inulin

Liu

Chi

et al. 2015

Mar Biotechnol

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In this study, in order to directly and efficiently convert inulin into pullulan, the INU1 gene from Kluyveromyces maximum KM was integrated into the genomic DNA and actively expressed in the high pullulan producer Aureobasidium melanogenum P16 isolated from the mangrove ecosystem. After the ability to produce pullulan from inulin by different transformants was examined, it was found that the recombinant strain EI36, one of the transformants, produced 40.92 U/ml of inulinase activity while its wild-type strain P16 only yielded 7.57 U/ml of inulinase activity. Most (99.27 %) of the inulinase produced by the recombinant strain EI36 was secreted into the culture. During the 10-l fermentation, 70.57 ± 1.3 g/l of pullulan in the fermented medium was attained from inulin (138.0 g/l) within 108 h, high inulinase activity (42.03 U/ml) was produced within 60 h, the added inulin was actively hydrolyzed by the secreted inulinase, and most of the reducing sugars were used by the recombinant strain EI36. This confirmed that the genetically engineered yeast of A. melanogenum strain P16 was suitable for direct pullulan production from inulin.

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“…This yeast has also been observed to readily secrete enzymes, an attribute that has been utilized for simultaneous saccharification and fermentation of cellulose for ethanol production (Ballesteros, Oliva, Negro, Manzanares, & Ballesteros, ; Tomás‐Pejó, Oliva, González, Ballesteros, & Ballesteros, ). These traits make K. marxianus particularly suited to industrial production of enzymes such as pectinase (Espinoza, Bárzana, García‐Garibay, & Ǵmez‐Ruiz, ), inulinase (Galindo‐Leva et al, ; Kushi, Monti, & Contiero, ), and β‐galactosidase (Rech, Cassini, Secchi, & Ayub, ; H. X. Zhou, Xu, Chi, Liu, & Chi, ), PCV2 virus‐like particles (Duan et al, ), as well as other chemicals such as ethyl acetate (Löbs, Engel, Schwartz, Flores, & Wheeldon, ; Löser, Urit, Gruner, & Bley, ), xylitol (Kim, Park, Jang, & Ha, ; Zhang et al, ), and 2‐phenylethanol (Gao & Daugulis, ; Martínez, Sánchez, Font, & Barrena, ).…”

Section: Introductionmentioning

confidence: 99%

Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

McTaggart

Bever

Bassett

et al. 2019

Biotech & Bioengineering

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Kluyveromyces marxianus is a promising nonconventional yeast for biobased chemical production due to its rapid growth rate, high TCA cycle flux, and tolerance to low pH and high temperature. Unlike Saccharomyces cerevisiae, K. marxianus grows on low-cost substrates to cell densities that equal or surpass densities in glucose, which can be beneficial for utilization of lignocellulosic biomass (xylose), biofuel production waste (glycerol), and whey (lactose). We have evaluated K. marxianus for the synthesis of polyketides, using triacetic acid lactone (TAL) as the product. The 2-pyrone synthase (2-PS) was expressed on a CEN/ARS plasmid in three different strains, and the effects of temperature, carbon source, and cultivation strategy on TAL levels were determined.The highest titer was obtained in defined 1% xylose medium at 37°C, with substantial titers at 41 and 43°C. The introduction of a high-stability 2-PS mutant and a promoter substitution increased titer four-fold. 2-PS expression from a multi-copy pKD1-based plasmid improved TAL titers a further five-fold. Combining the best plasmid, promoter, and strain resulted in a TAL titer of 1.24 g/L and a yield of 0.0295 mol TAL/mol carbon for this otherwise unengineered strain in 3 ml tube culture. This is an excellent titer and yield (on xylose) before metabolic engineering or fed-batch culture relative to other hosts (on glucose), and demonstrates the promise of this rapidly growing and thermotolerant yeast species for polyketide production.

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β-Galactosidase over-production by a mig1 mutant of Kluyveromyces marxianus KM for efficient hydrolysis of lactose

Cited by 21 publications

References 21 publications

Release of glucose repression on xylose utilization in Kluyveromyces marxianus to enhance glucose-xylose co-utilization and xylitol production from corncob hydrolysate

Release of glucose repression on xylose utilization in Kluyveromyces marxianus to enhance glucose-xylose co-utilization and xylitol production from corncob hydrolysate

Genetic Modification of the Marine-Isolated Yeast Aureobasidium melanogenum P16 for Efficient Pullulan Production from Inulin

Synthesis of polyketides from low cost substrates by the thermotolerant yeast Kluyveromyces marxianus

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