Eruca sativa is a rocket plant and a member of the Brassicaceae, which is considered to be an important chemo-preventive plant family. Although Eruca sativa has positive biological effects, the effect of Eruca sativa extract (ES) on improvement of skin barrier function has not been reported. In this study, we investigated the applicability of functional materials by examining a variety of physiological activities of Eruca sativa extract. ES showed anti-microbial activities against Bacillus subtilis, Escherichia coli, and Candida albicans. In particular, antimicrobial activities of ES against B. subtilis was the highest. Additionally, immunohistochemical analysis of protein marker related to keratinocyte differentiation was determined. The treatment by ES (50 mg/L) showed a significant increase of involucrin expression compared with treatment by 0.1% DMSO as a control in skin equivalents, the ES-treated group showed similar level in the expression of involucrin compared to the group treated with the same concentration of WY14643 in EpiDerm™, a three-dimensional model of skin equivalents. These results indicate that ES promotes the expression of protein related to barrier properties of the skin. Therefore, ES may be an effective ingredient for skin barrier improvement.
Lovastatin (also known as Mevinolin, Mevacor, and Monacolin K), an inhibitor of the HMG-CoA reductase produced by Aspergillus terreus and other fungi, is used to reduce serum cholesterol levels in human beings. It is derived biosynthetically from two polyketides. One of these is a nonaketide that undergoes cyclization at a hexahydronaphthalene ring system, and the other is a simple diketide, 2-methylbutyrate. Two primer pairs were designed based on the amino acid sequences of lovastatin polyketide synthase and lovastatin diketide synthase for the PCR screening of lovastatin-producing strains. Among the seven selected strains, J-2 evidenced the highest level of lovastatin production in both liquid and solid cultures. Soybeans with SJ-2 were treated via 1 hour of heat shock at 30℃ for the mass production of lovastatin. The heat-treated soybeans were inoculated on rice bran and the koji extract was obtained after 15 days of incubation. It yielded the highest level of lovastatin production among the strains, and also evidenced 75% inhibition activity against HMG-CoA reductase. We developed an efficient PCR screening method for lovastatin-producing strains, using lovastatin biosynthesis genes.
Peroxisome proliferator-activated receptors (PPAR)-α plays an important role in epidermal differentiation and barrier recovery, and topical treatment with PPAR-α agonists restores epidermal homeostasis in essential fatty acid deficiency and permeability barrier in skin disruptions. Therefore, we performed structure-based pharmacophore screening to search for a novel PPAR-α agonist. Caffeic acid was ultimately selected and evaluated for its effects on keratinocyte differentiation and epidermal permeability barrier.The transactivation activity of PPAR-responsive element (PPRE) and cornified envelope (CE) formation were assayed. Also, immunoblot analysis and anti-oxidant activity were investigated on caffeic acid.Caffeic acid increases the transactivation activity of PPRE and CE formation in keratinocytes. In addition, caffeic acid promotes the expression of genes and proteins related to CE formation such as involucrin and transglutaminase-1. Additionally, anti-oxidant activity were improved by caffeic acid.Caffeic acid can promote keratinocyte differentiation and restore skin barrier homeostasis and is suggested to be an appropriate skin therapeutic agent for improving epidermal permeability barrier function.
본 연구는 Asp. terreus ATCC 20542 변이주로부터 lovastatin 생산용 seed culture의 대량제조를 위한 방법을 개발한 것이다. 배양체의 선발, 분석 및 최적 배양용기를 검토한 결과 대두를 이용하여 petri dish(ø150×20 ㎜)에 배양하였을 때 lovastatin의 생산성이 우수하였다. 포자의 발아 촉진을 위하여 대두에 Asp. terreus를 접종한 다음 열처리를 달리하여, 각 전배양체를 미강에 본배양하였다. 본배양액을 추출한 후 HPLC를 이용하여 lovastatin 생산량을 검토한 결과 30℃에서 1시간 동안 열처리한 전배양체가 본배양 12일째에 가장 높은 lovastatin 생산성을 보이며, in vitro assay 결과, 대두를 30℃에서 1시간 열처리하여 본배양하였을 경우에 HMG-CoA reductase가 82% 저해되는 것으로 나타났다. 따라서 기존의 포자현탁액 접종법보다 대두를 이용한 방법이 더욱 높은 HMG-CoA reductase 저해활성 및 배양시간의 단축성을 보여 산업화에 유리한 것으로 사료되었다.
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