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  • 1
    Online-Ressource
    Online-Ressource
    Wiley ; 2019
    In:  MicrobiologyOpen Vol. 8, No. 9 ( 2019-09)
    In: MicrobiologyOpen, Wiley, Vol. 8, No. 9 ( 2019-09)
    Kurzfassung: A novel oligotrophic bacterium, designated strain CCA6, was isolated from leaf soil collected in Japan. Cells of the strain were found to be a Gram‐negative, non‐sporulating, motile, rod‐shaped bacterium. Strain CCA6 grew at 10–45°C (optimum 20°C) and pH 4.5–10.0 (optimum pH 5.0). The strain was capable of growth in poor‐nutrient (oligotrophic) medium, and growth was unaffected by high‐nutrient medium. The major fatty acid and predominant quinone system were C 16:0 and ubiquinone‐8. Phylogenetic analysis based on 16S rRNA gene sequences indicated strain CCA6 presented as a member of the family Enterobacteriaceae . Multilocus sequence analysis (MLSA) based on fragments of the atpD , gyrB , infB , and rpoB gene sequences was performed to further identify strain CCA6. The MLSA showed clear branching of strain CCA6 with respect to Enterobacter type strains. The complete genome of strain CCA6 consisted of 4,476,585 bp with a G+C content of 54.3% and comprising 4,372 predicted coding sequences. The genome average nucleotide identity values between strain CCA6 and the closest related Enterobacter type strain were 〈 88.02%. Based on its phenotypic, chemotaxonomic and phylogenetic features, strain CCA6 (=HUT 8142 T =KCTC 62525 T ) can be considered as a novel species within the genus Enterobacter with the proposed name Enterobacter oligotrophica .
    Materialart: Online-Ressource
    ISSN: 2045-8827 , 2045-8827
    URL: Issue
    Sprache: Englisch
    Verlag: Wiley
    Publikationsdatum: 2019
    ZDB Id: 2661368-2
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 2
    In: Journal of Mass Spectrometry, Wiley, Vol. 54, No. 3 ( 2019-03), p. 213-221
    Kurzfassung: A rapid analytical methodology is proposed to answer the two questions about the molecular and structural features of the acidic xylo‐oligosaccharides (XOSs) formed upon the enzymatic hydrolysis of 4‐ O ‐methylglucuronoxylan. The shortest acidic XOSs carrying a methylglucuronic acid moiety and the possible distribution of larger products (molecular feature) are instantly found by electrospray ionization mass spectrometry (ESI‐MS) in the negative ion mode, which filters the unwanted neutral XOS. The acidic moiety is then unambiguously localized along the xylose backbone (structural feature) by ESI‐MS n in the negative ion mode via the selection/activation/dissociation of the product ions formed upon the one‐way and stepwise glycosidic bond cleavage at the reducing end. Using the shortest acidic XOS with a known shape generated by glycoside hydrolase family (GH) 10 and GH11 xylanases as a proof of principle, pairs of diagnostic ions are proposed to instantly interpret the MS n fingerprints and localize the acidic moiety along the xylose chain of the activated ion. The original structure of the acidic XOS is then reconstructed by adding as many xylose units at the reducing end as MS n steps. Relying on pairs of ions, the methodology is robust enough to highlight the presence of isomeric products. Mass spectra reported in the present article will be conveniently used as reference data for the forthcoming analysis of acidic XOS generated by new classes of enzymes using this multistage mass spectrometry methodology.
    Materialart: Online-Ressource
    ISSN: 1076-5174 , 1096-9888
    URL: Issue
    Sprache: Englisch
    Verlag: Wiley
    Publikationsdatum: 2019
    ZDB Id: 2197367-2
    ZDB Id: 1472468-6
    ZDB Id: 7414-7
    SSG: 11
    SSG: 12
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 3
    In: MicrobiologyOpen, Wiley, Vol. 9, No. 8 ( 2020-08)
    Kurzfassung: meso ‐Diaminopimelate dehydrogenase ( meso ‐DAPDH) catalyzes the reversible NADP + ‐dependent oxidative deamination of meso ‐2,6‐diaminopimelate ( meso ‐DAP) to produce l ‐2‐amino‐6‐oxopimelate. Moreover, d ‐amino acid dehydrogenase ( d ‐AADHs) derived from protein‐engineered meso ‐DAPDH is useful for one‐step synthesis of d ‐amino acids with high optical purity. Here, we report the identification and functional characterization of a novel NAD(P) + ‐dependent meso ‐DAPDH from Numidum massiliense (NmDAPDH). After the gene encoding the putative NmDAPDH was expressed in recombinant Escherichia coli cells, the enzyme was purified 4.0‐fold to homogeneity from the crude extract through five purification steps. Although the previously known meso ‐DAPDHs use only NADP + as a coenzyme, NmDAPDH was able to use both NADP + and NAD + as coenzymes. When NADP + was used as a coenzyme, NmDAPDH exhibited an approximately 2 times higher k cat / K m value toward meso ‐DAP than that of meso ‐DAPDH from Symbiobacterium thermophilum (StDAPDH). NmDAPDH also catalyzed the reductive amination of corresponding 2‐oxo acids to produce acidic d ‐amino acids such as d ‐aspartate and d ‐glutamate. The optimum pH and temperature for the oxidative deamination of meso ‐DAP were about 10.5 and 75°C, respectively. Like StDAPDH, NmDAPDH exhibited high stability: it retained more than 75% of its activity after 30 min at 60°C (pH 7.2) or at pHs ranging from 5.5 to 13.0 (50°C). Alignment of the amino acid sequences of NmDAPDH and the known meso ‐DAPDHs suggested NmDAPDH has a hexameric structure. Given its specificity for both NADP + and NAD + , high stability, and a broad range of reductive amination activity toward 2‐oxo acids, NmDAPDH appears to offer advantages for engineering a more effective d ‐AADH.
    Materialart: Online-Ressource
    ISSN: 2045-8827 , 2045-8827
    URL: Issue
    Sprache: Englisch
    Verlag: Wiley
    Publikationsdatum: 2020
    ZDB Id: 2661368-2
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 4
    Online-Ressource
    Online-Ressource
    Wiley ; 2020
    In:  FEBS Open Bio Vol. 10, No. 6 ( 2020-06), p. 1180-1189
    In: FEBS Open Bio, Wiley, Vol. 10, No. 6 ( 2020-06), p. 1180-1189
    Kurzfassung: Xylanase B, a member of subfamily 7 of the GH30 (glycoside hydrolase family 30) from Talaromyces cellulolyticus ( Tc Xyn30B), is a bifunctional enzyme with glucuronoxylanase and xylobiohydrolase activities. In the present study, crystal structures of the native enzyme and the enzyme–product complex of Tc Xyn30B expressed in Pichia pastoris were determined at resolutions of 1.60 and 1.65 Å, respectively. The enzyme complexed with 2 2 ‐(4‐ O ‐methyl‐α‐ d ‐glucuronyl)‐xylobiose (U 4m2 X) revealed that Tc Xyn30B strictly recognizes both the C‐6 carboxyl group and the 4‐ O ‐methyl group of the 4‐ O ‐methyl‐α‐ d ‐glucuronyl side chain by the conserved residues in GH30‐7 endoxylanases. The crystal structure and site‐directed mutagenesis indicated that Asn‐93 on the β2‐α2‐loop interacts with the non‐reducing end of the xylose residue at subsite‐2 and is likely to be involved in xylobiohydrolase activity. These findings provide structural insight into the mechanisms of substrate recognition of GH30‐7 glucuronoxylanase and xylobiohydrolase.
    Materialart: Online-Ressource
    ISSN: 2211-5463 , 2211-5463
    URL: Issue
    Sprache: Englisch
    Verlag: Wiley
    Publikationsdatum: 2020
    ZDB Id: 2651702-4
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 5
    In: Biotechnology and Bioengineering, Wiley
    Kurzfassung: The integration of first‐ (1G) and second‐generation (2G) ethanol production by adding sugarcane juice or molasses to lignocellulosic hydrolysates offers the possibility to overcome the problem of inhibitors (acetic acid, furfural, hydroxymethylfurfural and phenolic compounds), and add nutrients (such as salts, sugars and nitrogen sources) to the fermentation medium, allowing the production of higher ethanol titers. In this work, an 1G2G production process was developed with hemicellulosic hydrolysate (HH) from a diluted sulfuric acid pretreatment of sugarcane bagasse and sugarcane molasses. The industrial Saccharomyces cerevisiae CAT‐1 was genetically modified for xylose consumption and used for co‐fermentation of sucrose, fructose, glucose, and xylose. The fed‐batch fermentation with high cell density that mimics an industrial fermentation was performed at bench scale fermenter, achieved high volumetric ethanol productivity of 1.59 g L −1 h −1 , 0.39 g g −1 of ethanol yield, and 44.5 g L −1 ethanol titer, and shown that the yeast was able to consume all the sugars present in must simultaneously. With the results, it was possible to establish a mass balance for the global process: from pretreatment to the co‐fermentation of molasses and HH, and it was possible to establish an effective integrated process (1G2G) with sugarcane molasses and HH co‐fermentation employing a recombinant yeast.
    Materialart: Online-Ressource
    ISSN: 0006-3592 , 1097-0290
    Sprache: Englisch
    Verlag: Wiley
    Publikationsdatum: 2024
    ZDB Id: 1480809-2
    ZDB Id: 280318-5
    SSG: 12
    Standort Signatur Einschränkungen Verfügbarkeit
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