• Toxicological Research
• /
• 제26권1호
• /
• pp.47-52
• /
• 2010

The Malassezia fungi are responsible for various human skin disorders including dandruff and seborrheic dermatitis. Of the Malassezia fungi, Malassezia globosa (M. globosa) is one of the most common in human scalp. The completed genome sequence of M. globosa contains four putative cytochrome P450 genes. To determine the roles of Malassezia P450 enzymes in the biosynthesis of ergosterol, we isolated MGL3996 gene from M. globosa chromosomal DNA by PCR. The MGL3996 gene encodes an enzyme of 616 amino acids, which shows strong similarity with known CYP52s of other species. MGL3996 gene was cloned and expressed in Pichia pastoris (P. pastoris) heterologous yeast expression system. Using the yeast microsomes expressing MGL3996 protein, a typical P450 CO-difference spectrum was shown with absorption maximum at 448 nm. SDS-PAGE analysis revealed a protein band of apparent molecular weight 69 kDa and Western blot with anti-histidine tag antibody showed that MGL3996 was successfully expressed in P. pastoris. Cloning and expression of a new P450 gene is an important step to study the P450 monooxygenase system of M. globosa and to understand the role of P450 enzymes in pathophysiology of dandruff.

• Journal of Microbiology and Biotechnology
• /
• 제22권1호
• /
• pp.141-146
• /
• 2012

Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

• Journal of Microbiology and Biotechnology
• /
• 제31권5호
• /
• pp.637-644
• /
• 2021

Malassezia is the most abundant genus in the fungal microflora found on human skin, and it is associated with various skin diseases. Among the 18 different species of Malassezia that have been identified to date, M. restricta and M. globosa are the most predominant fungal species found on human skin. Several studies have suggested a possible link between Malassezia and skin disorders. However, our knowledge on the physiology and pathogenesis of Malassezia in human body is still limited. Malassezia is unable to synthesize fatty acids; hence, it uptakes external fatty acids as a nutrient source for survival, a characteristic compensated by the secretion of lipases and degradation of sebum to produce and uptake external fatty acids. Although it has been reported that the activity of secreted lipases may contribute to pathogenesis of Malassezia, majority of the data were indirect evidences; therefore, enzymes' role in the pathogenesis of Malassezia infections is still largely unknown. This review focuses on the recent advances on Malassezia in the context of an emerging interest for lipases and summarizes the existing knowledge on Malassezia, diseases associated with the fungus, and the role of the reported lipases in its physiology and pathogenesis.

• 대한화장품학회지
• /
• 제38권4호
• /
• pp.311-319
• /
• 2012

보통 지루성 피부염은 과다 생성된 피지와 함께 피지를 영양분으로 상재하는 M. globosa 이스트에 의해서 얼굴, 몸 및 두피에 자주 발생하는 피부질환이다. 본 연구는 천연정유성분(Natural essential oils)으로부터 지루성 피부염과 관련된 항염증 활성과 M. globosa 이스트에 대한 증식 억제 활성을 평가하고자 하였다. 지루성 피부염 진정 효능을 평가하기 위해서 지루성 피부염 환경을 모사한 공배양 세포 평가 모델을 구축하였다. 본 평가모델을 통해서 육계(Cinnamomum zeylanicum, Cinnamon bark)의 껍질 및 박하(Mentha arvensis, Cornmint)의 잎에서 추출한 천연정유 성분이 다양한 지루성 피부염 자극원에 따른 Interleukin-$1{\alpha}$ (IL-$1{\alpha}$), Interleukin-6 (IL-6), Interleukin-8 (IL-8) 사이토카인의 발현량 증가와 피지생성 증가를 감소시킴을 확인하였다. 또한, 지루성 피부염 유발균으로 알려진 M. globosa 이스트에 대한 증식 저해 평가를 통해 두 정유성분에서 유의한 M. globosa 증식 억제 능(MIC ${\leq}0.0625%$)을 확인하였다. 본 연구 결과로부터 육계껍질 및 박하 잎 정유성분이 지루성 피부염에 따른 염증성 사이토카인의 발현량을 감소시켰다. 특히 육계 껍질 오일은 지루성 피부염 주요 원인인 Malassezia furfur 및 M. globosa 증식을 억제함으로써, 지루성 두피 염증의 예방과 효과적인 관리 제품의 소재로서 활용가치가 높음을 알 수 있다.

• Journal of Microbiology and Biotechnology
• /
• 제25권11호
• /
• pp.1827-1834
• /
• 2015

The SMG1 lipase from Malassezia globosa is a newly found mono- and diacylglycerol (DAG) lipase that has a unique lid in the loop conformation that differs from the common alpha-helix lid. In the present study, we characterized the contribution of three residues, L103 and F104 in the lid and F278 in the rim of the binding site groove, on the function of SMG1 lipase. Site-directed mutagenesis was conducted at these sites, and each of the mutants was expressed in the yeast Pichia pastoris, purified, and characterized for their activity toward DAG and p-nitrophenol (pNP) ester. Compared with wild-type SMG1, F278A retained approximately 78% of its activity toward DAG, but only 11% activity toward pNP octanoate (pNP-C8). L103G increased its activity on pNP-C8 by approximately 2-fold, whereas F104G showed an approximate 40% decrease in pNP-C8 activity, and they both showed decreased activity on the DAG emulsion. The deletion of 103-104 retained approximately 30% of its activity toward the DAG emulsion, with an almost complete loss of pNP-C8 activity. The deletion of 103-104 showed a weaker penetration ability to a soybean phosphocholine monolayer than wild-type SMG1. Based on the modulation of the specificity and activity observed, a pNP-C8 binding model for the ester (pNP-C8, N102, and F278 form a flexible bridge) and a specific lipid-anchoring mechanism for DAG (L103 and F104 serve as "anchors" to the lipid interface) were proposed.

• Journal of Microbiology and Biotechnology
• /
• 제19권8호
• /
• pp.743-748
• /
• 2009

Fungal diversity during composting was investigated by culture-independent rDNA sequence analysis. Composting was carried out with pig manure and mushroom cultural waste using a field-scale composter (Hazaka system), and samples were collected at various stages. Based on partial sequence analysis of large subunit (LSU) ribosomal RNA (rRNA) and sequence identity values, a total of 12 different fungal species were found at six sampling sites; Geotrichum sp., Debaryomyces hansenii, Monographella nivalis, Acremonium strictum, Acremonium alternatum, Cladosporium sphaerospermum, Myriangium durosai, Pleurotus eryngii, Malassezia globosa, Malassezia restricta, Rhodotorula glutinis, and Fusarium sporotrichioides. Geotrichum sp. of the class Saccharomycetes was the most predominant fungal species throughout the composting process (185 out of a total of 236 identified clones, or 78.4%), followed by Acremonium strictum (7.6%), Monographella nivalis (5.1%), and Pleurotus eryngii (3.8%). The prevalence of Geotrichum sp. was the lowest (61.1%) at the beginning of composting, and then gradually increased to 92.5% after 10 days of composting.