NAD(P)H-dependent aldose reductase from the xylose-fermenting yeast Pichia stipitis

Verduyn, Cornelis; Kleef, R. van; Jzn, J. Frank; Schreuder, Herman; Dijken, Johannes P. van; Scheffers, W. A.

doi:10.1007/bf00404545

Cited by 50 publications

(88 citation statements)

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“…This result is in accordance with the literature, in so far as the optimum pH for XR activity was 5.5 for Candida guilliermondii (Silva, et al, 1996), 6.0 for Pachysolen tannophylus (Dietzelmuller et al, 1984) and Pichia stipitis (Verduyn et al, 1985). However, the maximum XR activity diminished about 44% after submission to the extraction (Figure 1), probably due to the direct effect of the organic solvent and CTAB on the structure of the enzyme (Kopp, Schede, 2004).…”

Section: Resultssupporting

confidence: 92%

Characterization of xylose reductase extracted by CTAB-reversed micelles from Candida guilliermondii homogenate

Cortez

Pessoa

Felipe³

et al. 2006

Rev. Bras. Cienc. Farm.

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Section: Resultssupporting

confidence: 92%

Characterization of xylose reductase extracted by CTAB-reversed micelles from Candida guilliermondii homogenate

Cortez

Pessoa

Felipe³

et al. 2006

Rev. Bras. Cienc. Farm.

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“…Within family 2, the XRs from Pichia stipitis (AKR2B1) and Pachysolen tannophilus (AKR2B3) also exhibit clear dual co-substrate specificity [24,25]. There is conflicting information, however, regarding XR from Candida tropicalis (AKR2B4).…”

Section: Discussionmentioning

confidence: 99%

Structure of xylose reductase bound to NAD+ and the basis for single and dual co-substrate specificity in family 2 aldo-keto reductases

Kavanagh

Klimacek

Nidetzky

et al. 2003

Biochemical Journal

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Xylose reductase (XR; AKR2B5) is an unusual member of aldoketo reductase superfamily, because it is one of the few able to efficiently utilize both NADPH and NADH as co-substrates in converting xylose into xylitol. In order to better understand the basis for this dual specificity, we have determined the crystal structure of XR from the yeast Candida tenuis in complex with NAD + to 1.80 Å resolution (where 1 Å = 0.1 nm) with a crystallographic R-factor of 18.3 %. A comparison of the NAD + -and the previously determined NADP + -bound forms of XR reveals that XR has the ability to change the conformation of two loops. To accommodate both the presence and absence of the 2 -phosphate, the enzyme is able to adopt different conformations for several different side chains on these loops, including Asn 276 , which makes alternative hydrogen-bonding interactions with the adenosine ribose. Also critical is the presence of Glu 227 on a short rigid helix, which makes hydrogen bonds to both the 2 -and 3 -hydroxy groups of the adenosine ribose. In addition to changes in hydrogen-bonding of the adenosine, the ribose unmistakably adopts a 3 -endo conformation rather than the 2 -endo conformation seen in the NADP + -bound form. These results underscore the importance of tight adenosine binding for efficient use of either NADH or NADPH as a co-substrate in aldo-keto reductases. The dual specificity found in XR is also an important consideration in designing a high-flux xylose metabolic pathway, which may be improved with an enzyme specific for NADH.

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“…2-Mercaptoethanol (2-ME) is a reducing agent often included in enzymatic reactions to prevent oxidation of the sulfhydryl group and hence maintain enzymatic activities (Verduyn et al, 1985). As shown in Figure 3D, the addition of NP-40 (1%) and 2-ME (10 mmol L 1 ) during MNase digestion only slightly affected the laddering pattern of MNase digestion, compared with the control group.…”

Section: Optimization Of Mnase Digestion For Macmentioning

confidence: 99%

Enzymatic and chemical mapping of nucleosome distribution in purified micro- and macronuclei of the ciliated model organism, Tetrahymena thermophila

Chen

Gao

Liu

et al. 2016

Sci. China Life Sci.

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Genomic distribution of the nucleosome, the basic unit of chromatin, contains important epigenetic information. To map nucleosome distribution in structurally and functionally differentiated micronucleus (MIC) and macronucleus (MAC) of the ciliate Tetrahymena thermophila, we have purified MIC and MAC and performed micrococcal nuclease (MNase) digestion as well as hydroxyl radical cleavage. Different factors that may affect MNase digestion were examined, to optimize mono-nucleosome production. Mono-nucleosome purity was further improved by ultracentrifugation in a sucrose gradient. As MNase concentration increased, nucleosomal DNA sizes in MIC and MAC converged on 147 bp, as expected for the nucleosome core particle. Both MNase digestion and hydroxyl radical cleavage consistently showed a nucleosome repeat length of ~200 bp in MAC of Tetrahymena, supporting ~50 bp of linker DNA. Our work has systematically tested methods currently available for mapping nucleosome distribution in Tetrahymena, and provided a solid foundation for future epigenetic studies in this ciliated model organism.

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NAD(P)H-dependent aldose reductase from the xylose-fermenting yeast Pichia stipitis

Cited by 50 publications

References 1 publication

Characterization of xylose reductase extracted by CTAB-reversed micelles from Candida guilliermondii homogenate

Characterization of xylose reductase extracted by CTAB-reversed micelles from Candida guilliermondii homogenate

Structure of xylose reductase bound to NAD+ and the basis for single and dual co-substrate specificity in family 2 aldo-keto reductases

Enzymatic and chemical mapping of nucleosome distribution in purified micro- and macronuclei of the ciliated model organism, Tetrahymena thermophila

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