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  1. Yahaya RSR, Normi YM, Phang LY, Ahmad SA, Abdullah JO, Sabri S
    Appl Microbiol Biotechnol, 2021 May;105(10):3955-3969.
    PMID: 33937928 DOI: 10.1007/s00253-021-11321-y
    Keratinase is an important enzyme that can degrade recalcitrant keratinous wastes to form beneficial recyclable keratin hydrolysates. Keratinase is not only important as an alternative to reduce environmental pollution caused by chemical treatments of keratinous wastes, but it also has industrial significance. Currently, the bioproduction of keratinase from native keratinolytic host is considered low, and this hampers large-scale usage of the enzyme. Straightforward approaches of cloning and expression of recombinant keratinases from native keratinolytic host are employed to elevate the amount of keratinase produced. However, this is still insufficient to compensate for the lack of its large-scale production to meet the industrial demands. Hence, this review aimed to highlight the various sources of keratinase and the strategies to increase its production in native keratinolytic hosts. Molecular strategies to increase the production of recombinant keratinase such as plasmid selection, promoter engineering, chromosomal integration, signal peptide and propeptide engineering, codon optimization, and glycoengineering were also described. These mentioned strategies have been utilized in heterologous expression hosts, namely, Escherichia coli, Bacillus sp., and Pichia pastoris, as they are most widely used for the heterologous propagations of keratinases to further intensify the production of recombinant keratinases adapted to better suit the large-scale demand for them. KEY POINTS: • Molecular strategies to enhance keratinase production in heterologous hosts. • Construction of a prominent keratinolytic host from a native strain. • Patent analysis of keratinase production shows rapid high interest in molecular field.
    Matched MeSH terms: Saccharomycetales
  2. Lorrine OE, Rahman RNZRA, Joo Shun T, Salleh AB, Oslan SN
    Anal Biochem, 2023 May 01;668:115092.
    PMID: 36889624 DOI: 10.1016/j.ab.2023.115092
    In eukaryotes, serine proteases are cellular localized hydrolases reported to regulate essential biological reactions. Improved industrial applications of proteins are aided by prediction and analysis of their 3-dimensional structures (3D). A serine protease was identified from CTG-clade yeast Meyerozyma guilliermondii strain SO and its 3D structure as well as its catalytic attributes have not been fully understood yet, thus we seek to report on the catalytic mechanism of M. guilliermondii strain SO MgPRB1 using substrate PMSF via in silico docking as well as its stability by way of disulfide bonds formation. Herein, bioinformatics tools and techniques were used to predict, validate and analyze the possible changes of CUG ambiguity (if any) in strain SO using template PDB ID: 3F7O. Structural assessments confirmed the classic catalytic triad Asp305, His337, and Ser499. Superimposition of MgPRB1 and template 3F7O structures revealed the unlinked cysteine residues between Cys341, Cys440, Cys471 and Cys506 of MgPRB1 compared to template 3F7O with two disulfide bonds formation, which confers structural stability. In conclusion, serine protease structure from strain SO was successfully predicted and studies towards understanding at the molecular level may be undertaken for its potential applications in the degradation of peptide bonds.
    Matched MeSH terms: Saccharomycetales*
  3. Lachance MA, Bowles JM, Wiens F, Dobson J, Ewing CP
    Int J Syst Evol Microbiol, 2006 Oct;56(Pt 10):2489-2493.
    PMID: 17012584 DOI: 10.1099/ijs.0.64452-0
    A novel species, Metschnikowia orientalis sp. nov., is described for haploid, heterothallic yeasts isolated from nitidulid beetles sampled in flowers in Rarotonga in the Cook Islands, and the Cameron Highlands of Malaysia. As evidenced by analysis of D1/D2 large subunit rDNA sequences, the species is related to Candida hawaiiana, to which it is similar in growth responses. Cylindrical, conjugated asci and acicular ascospores of moderate size are formed. Rudimentary mating reactions were observed with Metschnikowia aberdeeniae and Metschnikowia continentalis, but not with C. hawaiiana. The type strain of M. orientalis is UWOPS 99-745.6(T) (h(+)) (=CBS 10331(T)=NRRL Y-27991(T)) and the designated allotype is UWOPS 05-269.1 (h(-)) (=CBS 10330=NRRL Y-27992).
    Matched MeSH terms: Saccharomycetales/classification*; Saccharomycetales/genetics; Saccharomycetales/isolation & purification; Saccharomycetales/physiology
  4. Freitas LFD, Barriga EJC, Barahona PP, Lachance MA, Rosa CA
    Int J Syst Evol Microbiol, 2013 Nov;63(Pt 11):4324-4329.
    PMID: 24014626 DOI: 10.1099/ijs.0.052282-0
    Twenty-four yeast strains were isolated from ephemeral flowers of Ipomoea spp. and Datura sp. and their associated insects in the Galápagos Archipelago, Ecuador, and from Ipomoea spp. and associated insects in the Cameron Highlands, Malaysia. Sequences of the D1/D2 domains of the large subunit rRNA gene indicated that these strains belong to a novel yeast species of the Kodamaea clade, although the formation of ascospores was not observed. The closest relative is Candida restingae. The human-mediated dispersion of this species by transpacific contacts in ancient times is suggested. The name Kodamaea transpacifica f.a., sp. nov. is proposed to accommodate these isolates. The type strain is CLQCA-24i-070(T) ( = CBS 12823(T) = NCYC 3852(T)); MycoBank number MB 803609.
    Matched MeSH terms: Saccharomycetales/classification*; Saccharomycetales/genetics; Saccharomycetales/isolation & purification
  5. Mahazar NH, Zakuan Z, Norhayati H, MeorHussin AS, Rukayadi Y
    Pak J Biol Sci, 2017;20(3):154-159.
    PMID: 29023007 DOI: 10.3923/pjbs.2017.154.159
    BACKGROUND AND OBJECTIVE: Inoculation of starter culture in cocoa bean fermentation produces consistent, predictable and high quality of fermented cocoa beans. It is important to produce healthy inoculum in cocoa bean fermentation for better fermented products. Inoculum could minimize the length of the lag phase in fermentation. The purpose of this study was to optimize the component of culture medium for the maximum cultivation of Candida sp. and Blastobotrys sp.

    MATERIALS AND METHODS: Molasses and yeast extract were chosen as medium composition and Response Surface Methodology (RSM) was then employed to optimize the molasses and yeast extract.

    RESULTS: Maximum growth of Candida sp. (7.63 log CFU mL-1) and Blastobotrys sp. (8.30 log CFU mL-1) were obtained from the fermentation. Optimum culture media for the growth of Candida sp., consist of 10% (w/v) molasses and 2% (w/v) yeast extract, while for Blastobotrys sp., were 1.94% (w/v) molasses and 2% (w/v) yeast extract.

    CONCLUSION: This study shows that culture medium consists of molasses and yeast extract were able to produce maximum growth of Candida sp. and Blastobotrys sp., as a starter culture for cocoa bean fermentation.

    Matched MeSH terms: Saccharomycetales/classification; Saccharomycetales/growth & development*; Saccharomycetales/metabolism
  6. de Jong AW, Dieleman C, Carbia M, Mohd Tap R, Hagen F
    J Clin Microbiol, 2021 03 19;59(4).
    PMID: 33536293 DOI: 10.1128/JCM.03220-20
    Non-albicans Candida species are emerging in the nosocomial environment, with the multidrug-resistant (MDR) species Candida auris being the most notorious example. Consequently, rapid and accurate species identification has become essential. The objective of this study was to evaluate five commercially available chromogenic media for the presumptive identification of C. auris Two novel chromogenic formulations, CHROMagar Candida Plus (CHROMagar) and HiCrome C. auris MDR selective agar (HiMedia), and three reference media, CandiSelect (Bio-Rad), CHROMagar Candida (CHROMagar), and Chromatic Candida (Liofilchem), were inoculated with a collection of 9 genetically diverse C. auris strains and 35 strains from closely related comparator species. After 48 h of incubation, the media were evaluated for their ability to detect and identify C. auris All media had the same limitations in the differentiation of the more common species Candida dubliniensis and Candida glabrata Only on CHROMagar Candida Plus did C. auris colonies develop a species-specific coloration. Nevertheless, the closely related pathogenic species Candida pseudohaemulonii and Candida vulturna developed a similar appearance as C. auris on this medium. CHROMagar Candida Plus was shown to be superior in the detection and identification of C. auris, with 100% inclusivity for C. auris compared to 0% and 33% for the reference media and HiCrome C. auris MDR selective agar, respectively. Although C. vulturna and C. pseudohaemulonii can cause false positives, CHROMagar Candida Plus was shown to be a valuable addition to the plethora of mostly molecular methods for C. auris detection and identification.
    Matched MeSH terms: Saccharomycetales
  7. Salleh AB, Baharuddin SM, Rahman RNZRA, Leow TC, Basri M, Oslan SN
    Microorganisms, 2020 Nov 06;8(11).
    PMID: 33171893 DOI: 10.3390/microorganisms8111738
    Screening for a new yeast as an alternative host is expected to solve the limitations in the present yeast expression system. A yeast sample which was isolated from the traditional food starter 'ragi' from Malaysia was identified to contain Meyerozyma guilliermondii strain SMB. This yeast-like fungus strain SMB was characterized to assess its suitability as an expression host. Lipase activity was absent in this host (when assayed at 30 °C and 70 °C) and Hygromycin B (50 μg/mL) was found to be its best selection marker. Then, the hyg gene (Hygromycin B) was used to replace the sh ble gene (Zeocin) expression cassette in a Komagataella phaffii expression vector (designated as pFLDhα). A gene encoding the mature thermostable lipase from Bacillus sp. L2 was cloned into pFLDhα, followed by transformation into strain SMB. The optimal expression of L2 lipase was achieved using YPTM (Yeast Extract-Peptone-Tryptic-Methanol) medium after 48 h with 0.5% (v/v) methanol induction, which was 3 times faster than another K. phaffii expression system. In conclusion, a new host-vector system was established as a platform to express L2 lipase under the regulation of PFLD1. It could also be promising to express other recombinant proteins without inducers.
    Matched MeSH terms: Saccharomycetales
  8. Lee JW, Ong EBB
    Front Cell Dev Biol, 2020;8:619126.
    PMID: 33511130 DOI: 10.3389/fcell.2020.619126
    Aging is a complex biological process that occurs in all living organisms. Aging is initiated by the gradual accumulation of biomolecular damage in cells leading to the loss of cellular function and ultimately death. Cellular senescence is one such pathway that leads to aging. The accumulation of nucleic acid damage and genetic alterations that activate permanent cell-cycle arrest triggers the process of senescence. Cellular senescence can result from telomere erosion and ribosomal DNA instability. In this review, we summarize the molecular mechanisms of telomere length homeostasis and ribosomal DNA stability, and describe how these mechanisms are linked to cellular senescence and longevity through lessons learned from budding yeast.
    Matched MeSH terms: Saccharomycetales
  9. Ejike UC, Chan CJ, Lim CSY, Lim RLH
    Appl Microbiol Biotechnol, 2021 Apr;105(7):2799-2813.
    PMID: 33763709 DOI: 10.1007/s00253-021-11225-x
    Fungal immunomodulatory proteins (FIPs) are bioactive proteins with immunomodulatory properties. We previously reported the heterologous production in Escherichia coli of FIP-Lrh from Tiger milk mushroom (Lignosus rhinocerus) with potent cytotoxic effect on cancer cell lines. However, protein produced in E. coli lacks post-translational modifications and may be contaminated with lipopolysaccharide (LPS) endotoxin. Therefore, in this study, yFIP-Lrh produced in Pichia pastoris was functionally compared with eFIP-Lrh produced in E. coli. Expression construct of FIP-Lrh cDNA in pPICZα was generated, transformed into P. pastoris X-33 and Mut+ transformants were verified by colony PCR. Induction with 0.5% or 1% methanol resulted in a secreted 13.6 kDa yFIP-Lrh which was subsequently purified and verified using LCMS/MS analysis. Size exclusion chromatography confirmed eFIP-Lrh as a homodimer whereas the larger size of yFIP-Lrh may indicate post-translational modification despite negative for glycoproteins staining. At lower concentration (4-8 μg/mL), yFIP-Lrh induced significantly higher Th1 (IFN-γ, TNF-α) and Th2 (IL-6, IL-4, IL-5, IL-13) cytokines production in mice splenocytes, whereas 16 μg/mL eFIP-Lrh induced significantly higher pro-inflammatory cytokines (TNF-α, IL-6, IL-10), possibly due to higher residual LPS endotoxin (0.082 EU/mL) in eFIP-Lrh compared to negligible level in yFIP-Lrh (0.001 EU/mL). Furthermore, yFIP-Lrh showed higher cytotoxic effect on MCF-7 and HeLa cancer cells. Since both recombinant proteins of FIP-Lrh have the same peptide sequence, besides glycosylation, other post-translational modifications in yFIP-Lrh may account for its enhanced immunomodulatory and anti-proliferative activities. In conclusion, P. pastoris is preferred over E. coli for production of a functionally active yFIP-Lrh devoid of endotoxin contamination. KEY POINTS: • FIP-Lrh can induced production of Th1 and Th2 cytokines by mouse splenocytes. • Higher cytotoxic effect on cancer cells observed for yeast compared to E. coli produced FIP-Lrh. • P. pastoris allows production of an endotoxin-free and functionally active recombinant FIP-Lrh.
    Matched MeSH terms: Saccharomycetales
  10. Ganapathy B, Yahya A, Ibrahim N
    Environ Sci Pollut Res Int, 2019 Apr;26(11):11113-11125.
    PMID: 30788704 DOI: 10.1007/s11356-019-04334-8
    Despite being a key Malaysian economic contributor, the oil palm industry generates a large quantity of environmental pollutant known as palm oil mill effluent (POME). Therefore, the need to remediate POME has drawn a mounting interest among environmental scientists. This study has pioneered the application of Meyerozyma guilliermondii with accession number (MH 374161) that was isolated indigenously in accessing its potential to degrade POME. This strain was able to treat POME in shake flask experiments under aerobic condition by utilising POME as a sole source of carbon. However, it has also been shown that the addition of suitable carbon and nitrogen sources has significantly improved the degradation potential of M. guilliermondii. The remediation of POME using this strain resulted in a substantial reduction of chemical oxygen demand (COD) of 72%, total nitrogen of 49.2% removal, ammonical nitrogen of 45.1% removal, total organic carbon of 46.6% removal, phosphate of 60.6% removal, and 92.4% removal of oil and grease after 7 days of treatment period. The strain also exhibited an extracellular lipase activity which promotes better wastewater treatment. Additionally, Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS) analyses have specifically shown that M. guilliermondii strain can degrade hydrocarbons, fatty acids, and phenolic compounds present in the POME. Ultimately, this study has demonstrated that M. guilliermondii which was isolated indigenously exhibits an excellent degrading ability. Therefore, this strain is suitable to be employed in the remediation of POME, contributing to a safe discharge of the effluent into the environment.
    Matched MeSH terms: Saccharomycetales/metabolism*
  11. Jinfeng EC, Mohamad Rafi MI, Chai Hoon K, Kok Lian H, Yoke Kqueen C
    World J Microbiol Biotechnol, 2017 Jan;33(1):5.
    PMID: 27844243
    Plants are primary source of natural product drugs. However, with every new bioactive molecule reported from a plant source, there follows reports of endangered status or even extinction of a medicinally important plant due to over-harvesting. Hence, the attention turned towards fungi namely the endophytes, which reside within medicinally important plants and thus may have acquired their medicinal properties. Strobilanthes crispus is a traditional medicinal plant which has been used traditionally to treat kidney stones, diabetes, hypertension and cancer as well as having antimicrobial activities. In our efforts to bioprospect for anticancer and antimicrobial metabolites, two fungal endophytes most closely related to the Sordariomycetes sp. showed promising results. Sample (PDA)BL3 showed highest significant antimicrobial activity against 6 bacteria at 200 µg/disc whereas sample (PDA)BL5 has highest significant anticancer activity against all 5 cancer cell lines at concentrations ranging from 30 to 300 μg/ml. As for the gas chromatography coupled with mass spectrometry (GC-MS) results, a total of 20 volatile metabolites identified from sample (PDA)BL3 and 21 volatile metabolites identified from sample (PDA)BL5 having more than 1% abundance. Both GC-MS analysis showed that compound Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl) has the highest abundance at 15.10% abundance for sample (PDA)BL3 and 19.00% abundance for sample (PDA)BL5 respectively. In conclusion, these results have shown bio-prospecting potential of endophytic fungi having antimicrobial and anticancer activities as well as its potential secondary metabolites of interest. Therefore, this work has further indicated the medicinal and industrial potential of endophytic fungi.
    Matched MeSH terms: Saccharomycetales/isolation & purification*; Saccharomycetales/chemistry
  12. Kurtzman CP
    Int J Syst Evol Microbiol, 2007 May;57(Pt 5):1154-1162.
    PMID: 17473275 DOI: 10.1099/ijs.0.64847-0
    The genus Blastobotrys, which now includes species previously assigned to the synonymous genera Arxula and Sympodiomyces, represents the anamorph of the ascosporogenous genus Trichomonascus. Six novel species are proposed for assignment to Blastobotrys. They were detected from their unique nucleotide sequences in large-subunit rDNA, ITS1-5.8S-ITS2 rDNA, mitochondrial small-subunit rDNA and the cytochrome oxidase II gene. The proposed novel species are Blastobotrys americana sp. nov. (type strain NRRL Y-6844(T)=CBS 10337(T); substrate unknown; Kansas, USA), Blastobotrys illinoisensis sp. nov. (type strain NRRL YB-1343(T)=CBS 10339(T); from forest debris; Illinois, USA), Blastobotrys malaysiensis sp. nov. (type strain NRRL Y-6417(T)=CBS 10336(T); from soil; Malaysia), Blastobotrys muscicola sp. nov. (type strain NRRL Y-7993(T)=CBS 10338(T); from moss; Louisiana, USA), Blastobotrys peoriensis sp. nov. (type strain NRRL YB-2290(T)=CBS 10340(T); from a fungus; Peoria, IL, USA) and Blastobotrys raffinosifermentans sp. nov. (type strain NRRL Y-27150(T)=CBS 6800(T); substrate unknown).
    Matched MeSH terms: Saccharomycetales/classification*; Saccharomycetales/cytology; Saccharomycetales/genetics
  13. Ding, C.H., Tzar M.N., Biswas S., Muttaqillah N.A.S., Wahab A.A.
    MyJurnal
    Catheter-related bloodstream infections caused by Kodamaea ohmeri are generally not considered due to the relative scarcity of reported cases. This is a case of an 85-year-old man with poorly controlled diabetes mellitus who was initially admitted to our hospital for diabetic ketoacidosis. An internal jugular catheter was inserted as part of the initial management. A week later the patient developed a temperature spike and a yeast identified as Kodamaea ohmeri by ID 32 C (bioMérieux, France) was isolated from both his central and peripheral blood cultures. The catheter was removed and the patient was treated with fluconazole despite the organism’s relatively high minimum inhibitory concentration (2 μg/mL) to this antifungal. The fungemia resolved following a 2-weeks course of fluconazole.
    Matched MeSH terms: Saccharomycetales
  14. Jacob AG, Wahab RA, Mahat NA
    Enzyme Microb Technol, 2021 Aug;148:109807.
    PMID: 34116744 DOI: 10.1016/j.enzmictec.2021.109807
    Oil palm leaves (OPL) silica (SiO2) can replace the energy-intensive, commercially produced SiO2. Moreover, the agronomically sourced biogenic SiO2 is more biocompatible and cost-effective enzyme support, which properties could be improved by the addition of magnetite (Fe3O4) and graphene oxide (GO) to yield better ternary support to immobilize enzymes, i.e., Candida rugosa lipase (CRL). This study aimed to optimize the Candida rugosa lipase (CRL immobilization onto the ternary OPL-silica-magnetite (Fe3O4)-GO (SiO2/Fe3O4/GO) support, for use as biocatalyst for ethyl valerate (EV) production. Notably, this is the first study detailing the CRL/SiO2/Fe3O4/GO biocatalyst preparation for rapid and high yield production of ethyl valerate (EV). AFM and FESEM micrographs revealed globules of CRL covalently bound to GL-A-SiO2/Fe3O4/GO; similar to Raman and UV-spectroscopy results. FTIR spectra revealed amide bonds at 3478 cm-1 and 1640 cm-1 from covalent interactions between CRL and GL-A-SiO2/Fe3O4/GO. Optimum immobilization conditions were 4% (v/v) glutaraldehyde, 8 mg/mL CRL, at 16 h stirring in 150 mM NaCl at 30 °C, offering 24.78 ± 0.26 mg/g protein (specific activity = 65.24 ± 0.88 U/g). The CRL/SiO2/Fe3O4/GO yielded 77.43 ± 1.04 % of EV compared to free CRL (48.75 ± 0.70 %), verifying the suitability of SiO2/Fe3O4/GO to hyperactivate and stabilize CRL for satisfactory EV production.
    Matched MeSH terms: Saccharomycetales
  15. Cheng Y, Lai OM, Tan CP, Panpipat W, Cheong LZ, Shen C
    ACS Appl Mater Interfaces, 2021 Jan 27;13(3):4146-4155.
    PMID: 33440928 DOI: 10.1021/acsami.0c17134
    Immobilization can be used to improve the stability of lipases and enhances lipase recovery and reusability, which increases its commercial value and industrial applications. Nevertheless, immobilization frequently causes conformational changes of the lipases, which decrease lipase catalytic activity. in the present work, we synthesized UIO-66 and grafted UIO-66 crystals with proline for immobilization of Candida rugosa lipase (CRL). As indicated by steady-state fluorescence microscopy, grafting of proline onto UIO-66 crystals induced beneficial conformational change in CRL. CRL immobilized on UIO-66/Pro (CRL@UIO-66/Pro) demonstrated higher enzyme activity and better recyclability than that immobilized on UIO-66 (CRL@UIO-66) in both hydrolysis (CRL@UIO-66/Pro: 0.34 U; CRL@UIO-66: 0.15 U) and transesterification (CRL@UIO-66/Pro: 0.93 U; CRL@UIO-66: 0.25 U) reactions. The higher values of kcat and kcat/Km of CRL@UIO-66/Pro also showed that it had better catalytic efficiency as compared to CRL@UIO-66. It is also worth noting that CRL@UIO-66/Pro (0.93 U) demonstrated a much higher transesterification activity as compared to free CRL (0.11 U), indicating that UIO-66/Pro has increased the solvent stability of CRL. Both CRL@UIO-66 and CRL@UIO-66/Pro were also used for the fabrication of biosensors for nitrofen with a wide linear range (0-100 μM), lower limit of detection, and good recovery rate.
    Matched MeSH terms: Saccharomycetales/enzymology*
  16. Jaafar NR, Littler D, Beddoe T, Rossjohn J, Illias RM, Mahadi NM, et al.
    Acta Crystallogr F Struct Biol Commun, 2016 11 01;72(Pt 11):831-839.
    PMID: 27827354
    Fuculose-1-phosphate aldolase (FucA) catalyses the reversible cleavage of L-fuculose 1-phosphate to dihydroxyacetone phosphate (DHAP) and L-lactaldehyde. This enzyme from mesophiles and thermophiles has been extensively studied; however, there is no report on this enzyme from a psychrophile. In this study, the gene encoding FucA from Glaciozyma antarctica PI12 (GaFucA) was cloned and the enzyme was overexpressed in Escherichia coli, purified and crystallized. The tetrameric structure of GaFucA was determined to 1.34 Å resolution. The overall architecture of GaFucA and its catalytically essential histidine triad are highly conserved among other fuculose aldolases. Comparisons of structural features between GaFucA and its mesophilic and thermophilic homologues revealed that the enzyme has typical psychrophilic attributes, indicated by the presence of a high number of nonpolar residues at the surface and a lower number of arginine residues.
    Matched MeSH terms: Saccharomycetales/enzymology; Saccharomycetales/chemistry*
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