• 대한의생명과학회지
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• 제20권4호
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• pp.244-249
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• 2014

Arazyme is a metalloprotease secreted by Aranicola proteolyticus that was previously shown to suppress cytokine expression of keratinocytes and endothelial cells and inhibit histopathological features in an atopic dermatitis-like animal model. However, the regulatory effects of arazyme in other allergic diseases have yet to be elucidated. In this study, we investigated whether arazyme is effective against neutrophil apoptosis in allergic diseases such as allergic rhinitis and asthma. Arazyme inhibited neutrophil apoptosis of normal subjects in a dose-dependent manner. However, the antiapoptotic effect of arazyme was reversed by LY294002, an inhibitor of PI3K, AKTi, an inhibitor of Akt, PD98059, an inhibitor of MEK, and BAY-11-7085, an inhibitor of NF-${\kappa}B$. Arazyme induced activation of NF-${\kappa}B$ via PI3K/Akt/ERK pathway. The anti-apoptotic effect of arazyme is associated with inhibition of cleavage of caspase 3 and caspase 9. Arazyme inhibited constitutive apoptosis of neutrophil in a dose-dependent manner in allergic subjects, and its mechanism was shown to be associated with PI3K/Akt/ERK/NF-${\kappa}B$. The results presented here improve our understanding of neutrophil apoptosis regulation and will facilitate development of drugs for treatment of allergic diseases.

• International Journal of Industrial Entomology and Biomaterials
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• 제48권2호
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• pp.67-77
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• 2024

This study aims to assess the efficacies of exogenous enzymes, derived from invertebrate gut-associated microbes, as feed additives, in reducing volatile organic compound (VOC) emissions using an in vitro pig intestine continuous fermentation system. An in vitro continuous fermentation model was used to simulate a comparable bionic digestion system by co-reacting feed, enzymatic additives (arazyme, mannanase, and xylanase, derived from the gut bacteria of Nephila clavata, Eisenia fetida, and Moechotypa diphysis, respectively), and gastrointestinal microbes, followed by an analysis of their correlations. A significant correlation was observed between exogenous enzyme supplementation and reduced VOC emissions in the fecal phase of continuous fermentation (p < 0.05). The concentration of VOCs decreased by 3.75 and 2.75 ppm in the treatment group following arazyme and multi-enzyme supplementation, respectively, compared to that in the control group (7.83 ppm). In addition, supplementation with arazyme and multiple enzymes significantly affected the microbial composition of each fermentation phase (p < 0.05). In particular, Lactiplantibacillus pentosus and Pediococcus pentosaceus, which changed in abundance according to arazyme or multi-enzyme supplementation, exhibited a positive relationship with VOC emissions. These results suggest that exogenous enzymes derived from invertebrate gut-associated bacteria can be efficiently applied as feed additives, leading to a reduction in VOC emissions.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.761-768
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• 2007

Serratia proteamaculans HY-3 isolated from the digestive tract of a spider produces an extracellular protease named arazyme, with an estimated molecular mass of 51.5 kDa. The purified enzyme was characterized as having high activities at wide pH and temperature ranges. We further characterized biochemical features of the enzymatic reactions under various reaction conditions. The protease efficiently hydrolyzed a broad range of protein substrates including albumin, keratin, and collagen. The dependence of enzymatic activities on the presence of metal ions such as calcium and zinc indicated that the enzyme is a metalloprotease, together with the previous observation that the proteolytic activity of the enzyme was not inhibited by aspartate, cysteine, or serine protease inhibitors, but strongly inhibited by 1,10-phenanthroline and EDTA. The araA gene encoding the exoprotease was isolated as a 5.6 kb BamHI fragment after PCR amplification using degenerate primers and subsequent Southern hybridization. The nucleotide sequence revealed that the deduced amino acid sequences shared extensive similarity with those of the serralysin family of metalloproteases from other enteric bacteria. A gene(inh) encoding a putative protease inhibitor was also identified immediately adjacent to the araA structural gene.

• 한국가금학회지
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• 제36권1호
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• pp.47-55
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• 2009

본 실험은 산란 후기 사료 내 거미 유래 단백질 분해효소 제 AZ의 첨가 급여가 산란계의 생산성, 계란의 품질, 혈액 특성, 장 내용물의 점도, 맹장 미생물 변화 및 맹장 내 암모니아 농도에 미치는 영향을 조사하기 위한 목적으로 수행하였다. 59주령의 Hy-Line Brown 실용계 200수를 공시하여 총 4개 처리에 각 처리구별 반복당 10수씩 5반복으로 완전 임의 배치하였고, 각각의 실험 사료에 AZ을 각각 0.05%, 0.075% 및 0.1% 혼합하여 총 5주간 급여하였다. 총 실험 기간 중 AZ 첨가에 따라 난중에서 유의한 차가 나타났으며, AZ 0.05%를 첨가한 처리구에서 가장 높은 증가가 나타났다(P<0.05). AZ 첨가는 난각 두께, 난각 강도, 난각색, 난황색 및 Haugh Unit는 처리구간 통계적 유의성은 없는 것으로 나타났다. 혈액 내 총 콜레스테롤 함량과 GOT 및 GPT 활성에 있어서는 처리구간 통계적 차이가 없는 것으로 나타났다. 맹장 내 미생물 균총을 조사해 본 결과 총 균수, 유산균수 및 대장균군수는 처리구간 차이를 보이지 않았다. 장 내용물의 점도 및 장내 암모니아 농도는 대조구에 비해 AZ을 첨가한 처리구에서 유의하게 감소하는 것으로 나타났다(P<0.05). 본 실험 결과, 식물성 및 동물성 단백질을 포함한 산란 후기 사료에 AZ의 첨가 급여는 난중 및 장 내 점도를 개선하고 암모니아 농도를 저감하는데 효과적이라는 것을 시사한다. 따라서 산란 후기 사료에 거미 유래 단백질 분해효소제 AZ의 첨가 급여는 산란 후기의 생산성, 장내 점도를 개선하여 사료 효소제로서 가치가 있는 것으로 판단된다. 또한 본 실험 결과 산란 후기 사료에서의 AZ의 첨가 수준은 0.05%와 0.075%에서 개선되는 결과가 나타났으나, 첨가 비용을 고려 할 때, 0.05% 첨가 수준이 가장 높은 효과가 있는 것으로 판단된다.