• 제목/요약/키워드: protein based carbon

검색결과 74건 처리시간 0.029초

Genomic and Proteomic Analysis of Microbial Function in the Gastrointestinal Tract of Ruminants - Review -

  • White, Bryan A.;Morrison, Mark
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권6호
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    • pp.880-884
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    • 2001
  • Rumen microbiology research has undergone several evolutionary steps: the isolation and nutritional characterization of readily cultivated microbes; followed by the cloning and sequence analysis of individual genes relevant to key digestive processes; through to the use of small subunit ribosomal RNA (SSU rRNA) sequences for a cultivation-independent examination of microbial diversity. Our knowledge of rumen microbiology has expanded as a result, but the translation of this information into productive alterations of ruminal function has been rather limited. For instance, the cloning and characterization of cellulase genes in Escherichia coli has yielded some valuable information about this complex enzyme system in ruminal bacteria. SSU rRNA analyses have also confirmed that a considerable amount of the microbial diversity in the rumen is not represented in existing culture collections. However, we still have little idea of whether the key, and potentially rate-limiting, gene products and (or) microbial interactions have been identified. Technologies allowing high throughput nucleotide and protein sequence analysis have led to the emergence of two new fields of investigation, genomics and proteomics. Both disciplines can be further subdivided into functional and comparative lines of investigation. The massive accumulation of microbial DNA and protein sequence data, including complete genome sequences, is revolutionizing the way we examine microbial physiology and diversity. We describe here some examples of our use of genomics- and proteomics-based methods, to analyze the cellulase system of Ruminococcus flavefaciens FD-1 and explore the genome of Ruminococcus albus 8. At Illinois, we are using bacterial artificial chromosome (BAC) vectors to create libraries containing large (>75 kbases), contiguous segments of DNA from R. flavefaciens FD-1. Considering that every bacterium is not a candidate for whole genome sequencing, BAC libraries offer an attractive, alternative method to perform physical and functional analyses of a bacterium's genome. Our first plan is to use these BAC clones to determine whether or not cellulases and accessory genes in R. flavefaciens exist in clusters of orthologous genes (COGs). Proteomics is also being used to complement the BAC library/DNA sequencing approach. Proteins differentially expressed in response to carbon source are being identified by 2-D SDS-PAGE, followed by in-gel-digests and peptide mass mapping by MALDI-TOF Mass Spectrometry, as well as peptide sequencing by Edman degradation. At Ohio State, we have used a combination of functional proteomics, mutational analysis and differential display RT-PCR to obtain evidence suggesting that in addition to a cellulosome-like mechanism, R. albus 8 possesses other mechanisms for adhesion to plant surfaces. Genome walking on either side of these differentially expressed transcripts has also resulted in two interesting observations: i) a relatively large number of genes with no matches in the current databases and; ii) the identification of genes with a high level of sequence identity to those identified, until now, in the archaebacteria. Genomics and proteomics will also accelerate our understanding of microbial interactions, and allow a greater degree of in situ analyses in the future. The challenge is to utilize genomics and proteomics to improve our fundamental understanding of microbial physiology, diversity and ecology, and overcome constraints to ruminal function.

Construction of fat1 Gene Expression Vector and Its Catalysis Efficiency in Bovine Fetal Fibroblast Cells

  • Liu, Boyang;Yang, Runjun;Li, Junya;Zhang, Lupei;Liu, Jing;Lu, Chunyan;Lian, Chuanjiang;Li, Zezhong;Zhang, Yong-Hong;Zhang, Liying;Zhao, Zhihui
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권5호
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    • pp.621-628
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    • 2012
  • The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

인류의 생존 위기 대응을 위한 양잠과 곤충 산업의 현황 (Current Status of Sericulture and Insect Industry to Respond to Human Survival Crisis)

  • 김아영;김기영;최희정;박현우;고영호
    • 한국응용곤충학회지
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    • 제61권4호
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    • pp.605-614
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    • 2022
  • 인류는 기후변화와 인구 구성 비율의 급격한 노령화 라는 두 가지 커다란 생존을 위협하는 문제점에 직면해 있다. 기후변화는 경제 발전과 운송 수단의 발달로 화석 연료 사용 증가에 따른 대기 중 온실가스 농도가 증가한 결과이고, 인구 구성 비율의 노령화는 선진국의 의 생명과학 기술 발전과 개인 위생의 증진으로 기대 수명이 증가한 결과이다. 돌이킬 수 없는 전 지구적인 기후변화를 피하기 위해서는 빠른 기간 내에 온실 가스의 배출이 없는 탄소 제로 경제로 전환을 해야 한다. 이 목표를 달성하기 위해서는 농업 중 온실가스의 발생이 가장 많은 낙농축산업을 저탄소 경영방식으로 전환하고 동시에 소비자들의 저탄소 식품들에 대한 인식의 변화가 필요하다. 현재 지구상 이용 가능 초지 중 77%가 가축용 사료 재배에 활용되지만, 인간이 섭취하는 전체 단백질의 37%와 총 열량의 18%만이 낙농축산업에서 얻어질 뿐이다. 그러므로, 가축보다 온실가스 배출량, 물의 사용량이 적고, 사육 공간이 작아도 되며 사료전환율이 높은 식용 곤충을 단백질원으로 활용해야 할 필요성이 있다. 이와 더불어 건강기능 증진 효과가 있다고 과학적으로 밝혀진 누에와 같은 곤충들의 기능성을 활용하여 현재 치료 방법이나 예방법이 확립되지 않은 퇴행성 질환들을 예방하고 치료를 촉진시킬 수 있는 기능성 식품 개발이 필요하다. 곤충은 동물 중 가장 오래 전에 지구상에 나타났고, 인간의 생존 유무와 상관없이 앞으로도 빠르게 진화를 하여 지구의 환경 변화에 적응하고 번성할 것이다. 그러므로, 다양한 식용 곤충과 누에를 포함한 약용 곤충을 이용한 산업은 현재 인류가 직면한 문제를 해결하여 미래에 인간이 지구에서 생존하고 번영할 수 있는 중요한 받침돌이 될 것이다.

양하분말을 이용한 녹두묵 제조 및 품질 평가 (Quality Characteristics of Nokdumook using Yangha(Zingiber mioga R.) Powder)

  • 김현순;김민정;이명숙;이건순;김애정
    • 한국식품영양학회지
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    • 제25권3호
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    • pp.521-528
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    • 2012
  • This study was performed to develop a therapeutic diet against aging and obesity, using Yangha(Zingiber mioga R.). Before development of a therapeutic diet, we performed cell viability assay, analysis of general composition, macrominerals and antioxidantive activities of Yangha. Based on the findings from analyzing the results, mook using Yangha powder(0~20%) was processed, and tested for quality characteristics such as color values, sensory evaluation and mechanical properties. The result of cell viability assay of myoga, using liver cells, revealed that within the concentration range from 500 ${\mu}g/m{\ell}$ to 10,000 ${\mu}g/m{\ell}$, cell survivability increased in line with the concentration rate. Therefore, it will not be harmful to consume it as food. Regarding the normal substance of myoga, the water substance of myoga was 94%, which exceeds that of ginger and tumeric with 89% and 83%, respectively. As for crude protein, fat, carbon hydrates and ash, myoga contained less than the other two, which I think is due to the high water substance. Regarding the minerals, potassium had the highest contents among macrominerals of 234.74 mg%. As for the antioxidant test, hydroxy radical scavenging activity and superoxide radical scavenging activity were shown. As for the production of Yangha mook(Yangha powder levels were 0~20%) for quality characteristics, the more of the powder, the less the L, but the greater the a and b values. Also, for the material property, an increased amount of the powder, resulted in chewiness and springiness, but less gumminess in a correlated manner. However, there were no significant differences in the springiness and cohesiveness in relation to the powder. For the sensory test, jelly type ZM5 with 5% powder showed highest overall preference. According to the sensory test, based on the powder substance, the jelly with 5% powder showed the highest overall score, including preference.

토양으로부터 새로이 분리된 단백질 분해효소 생산 미생물 Bacillus subtilis FBL-1의 동정 (Identification of a Newly Isolated Protease-producing Bacterium, Bacillus subtilis FBL-1, from Soil)

  • 김민아;시진범;위영중
    • 한국미생물·생명공학회지
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    • 제44권2호
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    • pp.185-193
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    • 2016
  • 경상북도 경산시에 위치한 영남대학교 내의 토양으로부터 protease를 생산하는 신규 미생물 FBL-1을 분리하였다. 본 균주는 Biolog test 및 API 50CHB test 결과, Bacillus 속 미생물 중 하나인 것으로 확인되었다. 16S rDNA 염기서열 분석결과로부터 FBL-1 균주는 B. subtilis subsp. subtilis NCIB 3610 (99.5%), B. subtilis subsp. inaquosorum KCTC 13429 (99.4%), B. subtilis subsp. spizizenii NRRL B-23049 (99.3%), B. methylotrophicus KACC 13105 (99.3%) 등과 높은 상동성을 보였다. 이러한 생리적, 형태학적, 계통학적 특성에 따라 균주 FBL-1은 B. subtilis에 속하는 균으로 최종 동정하여 B. subtilis FBL-1으로 명명하였다. B. subtilis FBL-1을 이용한 protease 생산시 탄소원으로는 fructose, 질소원으로는 yeast extract가 균체 성장 및 protease 활성을 위하여 가장 적합한 것으로 나타났다. B. subtilis FBL-1 균주는 protease를 생산하는데 활용할 수 있으며, 향후 배지 및 발효조건 최적화와 효소의 특성 규명 등의 연구를 통하여 식품, 세제 등의 다양한 산업에 유용하게 사용될 수 있을 것으로 기대된다.

경주 동궁과 월지 3호 우물 출토 옛사람 뼈의 동위원소에 기록된 고려시대 식생활 양상 (Isotopic palaeodiet studies of human bone from Gyeongju Donggung Palace and Wolji pond site (pond No.3), Goryeo period)

  • 최현구;신지영
    • 분석과학
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    • 제32권6호
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    • pp.262-270
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    • 2019
  • 뼈에서 추출된 콜라겐의 탄소와 질소 안정동위원소 정보는 그 당시의 사람들이 섭취했던 식생활, 영양 상태, 고환경과 생활상을 연구하는데 중요한 역할을 한다. 본 연구에서는 출토 자료가 희박하여 아직까지 시도된 바가 없는 고려시대 피장자들에 대한 식생활과 모유수유 양상을 밝히고자 한다. 고려시대 경주 동궁과 월지 3호 우물에서 출토된 옛사람 뼈 4개체에서 추출된 뼈 콜라겐을 이용하여 안정동위원소 분석을 진행하였으며, 탄소와 질소 안정동위원소 분석 결과는 각각 다음과 같다(δ13C(‰) = -19.5 ± 0.9 ‰, δ15N(‰) = 11.1 ± 1.1 ‰, (n = 4)). 탄소 안정동위원소 분석 결과 벼, 보리 등 C3 작물군을 주로 섭취하였고, 질소 안정동위원소 분석 결과 동물성 단백질은 주로 육상 동물로부터 얻었을 가능성이 높다고 추정된다. 또한, 출토된 옛사람 뼈 4개체는 성인, 소아, 유아, 영아로 추정되는 다양한 연령대로 나타났으며, 유아와 영아로 추정되는 피장자들과 성인 피장자 간의 질소 안정동위원소 값 차이로부터 약 3세의 연령까지 모유수유가 진행되고 있었다고 추정된다. 모유수유와 이유 양상은 당시의 출산율, 인구역학, 이동 패턴, 질병 등 다양한 사회상을 반영하는 매우 중요한 정보이며, 다양한 연령대의 분석 결과로부터 의미 있는 정보를 밝혀냈다.

Emissions of Odor, Ammonia, Hydrogen Sulfide, and Volatile Organic Compounds from Shallow-Pit Pig Nursery Rooms

  • Kafle, Gopi Krishna;Chen, Lide
    • Journal of Biosystems Engineering
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    • 제39권2호
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    • pp.76-86
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    • 2014
  • Purpose: The objective of this study was to measure emissions of gases (ammonia ($NH_3$), hydrogen sulfide ($H_2S$) and carbon dioxide ($CO_2$)), volatile organic compounds (VOC) and odor from two shallow pit pig nursery rooms. Gas and odor reduction practices for swine operations based on the literature were also discussed. Methods: This study was conducted for 60 days at a commercial swine nursery facility which consisted of four identical rooms with mechanical ventilations. Two rooms (room 1 (R1) and room 2 (R2)) with different pig numbers and ventilation rates were used in this study. The pig manure from both the R1 and R2 were characterized. Indoor/outdoor temperatures, ventilation rates/duration, $NH_3$, $H_2S$, $CO_2$, and VOC concentrations of the ventilation air were measured periodically (3-5 times/week). Odor concentrations of the ventilations were measured two times on two days. Three different types of gas and odor reduction practices (diet control, chemical method, and biological method) were discussed in this study. Results: The volatile solids to total solids ratio (VS/TS) and crude protein (CP) value of pig manure indicated the pig manure had high potential for gas and odor emissions. The $NH_3$, $H_2S$, $CO_2$ and VOC concentrations were measured in the ranges of 1.0-13.3, 0.1-5.7, 1600-3000 and 0.0-1.83 ppm, respectively. The $NH_3$ concentrations were found significantly higher than $H_2S$ concentrations for both rooms. The odor concentrations were measured in the range of $2853-4432OU_E/m^3$. There was significant difference in odor concentrations between the two rooms which was due to difference in pig numbers and ventilation duration. The literature studies showed that simultaneous use of dietary control and biofiltration practices will be more effective and environmentally friendly for gas and odor reductions from pig barns. Conclusions: The gas and odor concentrations measured in the ventilation air from the pig rooms indicate an acute need for using gas and odor mitigation technologies. Adopting diet control and biofiltration practices simultaneously could be the best option for mitigating gas and odor emissions from pig barns.

느타리 톱밥종균 배지조성에 따른 생산성 비교 (Comparison in the productivity of Pleurotus ostreatus sawdust spawn based on the medium composition)

  • 최종인;최준영;김정한;구옥;김연진;하태문;정구현
    • 한국버섯학회지
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    • 제19권1호
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    • pp.9-13
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    • 2021
  • 느타리 종균배지재료인 미송톱밥과 미강을 대체할 수 있는 재료를 선발하기 위하여 실시하였다. 톱밥의 pH는 4.9~6.1, 총질소함량은 0.21~0.26%, 총탄소함량은 49.0~50.2%를 나타내며 조지방은 0.3~0.7%를 함유하고 있다. 종균 혼합배지의 pH는 5.3~5.9, 총질소함량은 0.65~1.11%, 총탄소함량은 47.0~49.1%를 나타내었다. C/N율은 총질소 함량이 낮은 미루나무톱밥 혼합배지에서 높게 나타났으며, 단백질 함량은 미송톱밥이 함유되어 있는 배지에서 높게 나타났다. 느타리 균사생장은 미강보다 밀기울이 포함되어 있는 배지에서 균사생장 속도가 빨랐다. 수량은 '곤지7호', '춘추2호', '흑타리', '수한1호' 순으로 높았으며, T4(미루나무톱밥:참나무톱밥:미강) 배지에서 가장 높은 생산성을 나타내었으나 처리 간 유의성이 없었다. 느타리 농가에서는 자가종균 제조 시 주변에서 손쉽게 구할 수 있는 미루나무톱밥, 참나무톱밥을 단용 및 혼용으로 이용할 수 있으며, 보관중에 품질유지가 어려운 미강 대신에 밀기울을 사용하여 종균을 제조할 수 있을 것으로 판단된다.

한국(韓國) 재래산양(在來山羊)의 질소대사(窒素代謝) 및 기초대사량(基礎代謝量)에 관(關)한 연구(硏究) (Studies on the Nitrogenous Utilization and Basal Metabolism of Korean Native Goat)

  • 오홍록
    • 농업과학연구
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    • 제9권2호
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    • pp.546-555
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    • 1982
  • 한국재래산양(韓國在來山羊)에 대한 단백질(蛋白質)의 소화(消化), 흡수(吸收) 및 질소대사량(窒素代謝量)과 energy의 발생량(發生量)을 조사(調査)하기 위해서 생후(生後) 10개월령(個月令)인 생체중(生體重) 20kg의 모산양(牡山羊)과 생후(生後) 30개월령(個月令)인 생체중(生體重) 20kg의 빈산양(牝山羊)을 공시동물(供試動物)로 하고, 개방식(開放式) 중량측정(重量測定) 호흡시험장치(呼吸試驗裝置)를 이용(利用)하여, 사료(飼料)의 급여수준(給與水準)에 의(依)한 질소(窒素) 및 energy의 대사량(代謝量)을 측정(測定)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 재래산양(在來山羊)에 급여(給與)한 사료(飼料)는 품질(品質)이 중간정도(中間程度)의 orchard grass 건초(乾草)였으며, 1(일)日 평균(平均) 생체중(生體重)에 대(對)하여 0.66~0.92% 섭취(攝取)하였는데, 건초(乾草)의 품질(品質)이 우수(優秀)하여도 그 이상(以上)은 섭취(攝取)하지 못하였으며, 유산양(乳山羊)이나 면양(緬羊)에 비(比)하여 떨어지는데 이는 동물상내(動物箱內)에서 경경(璟境)의 변화(變化)에도 기인(基因)되는 것 같다. 농후사료(濃厚飼料)로 대두박(大豆粕)을 급여(給與)한 구(區)에서는 건초(乾草)를 1.06%, 대두박(大豆粕)을 0.6% 섭취하여 생체중(生體重)의 1.66%를 섭취하였다. 2. 개방식(開放式) 중량측정호흡법(重量測定呼吸法)에 의(依)한 $CO_2$깨스발생량(發生量)을 KOH 용액(溶液)에 흡수(吸收)시켜, 24시간(時間)의 증가량(增加量)을 측정(測定)한 결과(結果)는 99~117g이었으나, 사료섭취량(飼料攝取量)의 증가(增加)에 의(依)하여 $CO_2$gas 발생량(發生量)이 증가(增加)하는 경향(傾向)은 없으며, 체중(體重)의 증가와 $CO_2$gas 발생량(發生量)은 비례(比例)하였다. 3. Orchard grass만을 급여(給與)하였을때 단백질(蛋白質)의 소화율(消化率)은 24.0~41%였으며, 동건초(同乾草)와 대두박(大豆粕)을 급여(給與)하였을때는 58.2%였는데, orchard grass 건초(乾草)212g과 대두박(大豆粕)150g을 섭취하여 11.538g의 질소를 1일(日) 섭취하여도 질소축적(窒素蓄積)은 0.16g이었다. 이는 조단백질(粗蛋白質)로 환산(換算)하여 1(일)日 77.9g의 섭취가 되며, 생체중(生體重) 20kg내외(內外)의 재래산양(在來山羊)에 필요(必要)한 40~50g보다 많았으나, N.E의 발생량이 537Kcal였으므로, 단백질(蛋白質)이 energy로 분해(分解) 이용(利用)되었다. 4. Energy의 출납(出納)은 생후(生後) 10개월령(個月令)의 생체중(生體重) 20.5kg의 모산양(牡山羊)에게 orchard 건초(乾草) 158g을 급여(給與)한 구(區)에서 G.E로 624Kcal 섭취하였고 D.E 량(量)은 260Kcal, 급열발생량(給熱發生量)은 338Kcal로 78Kcal가 부족(不足)하였다. 또, 생체중(生體重) 20kg의 생후(生後)30개월령(個月令) 모산양(牡山羊)에게 건초(乾草)213g을 급여(給與)아여 G.E로 842Kcal를 섭취하였을때 D.E는 199Kcal였고, 총열발생량(총熱發生量)은 334Kcal로 135Kca1가 부족(不足)하였다. 동모산양(同牡山羊)에게 건초(乾草) 212g과 대두분(大豆粉)150g을 급여(給與)하여 G.E로 1609Kcal를 섭취하였을때 역시 54Kcal가 부족(不足)하였다. 따라서 생체중(生體重) 20kg 내외(內外)의 한국재래산양(韓國在來山羊)은 N.E로 600Kcal 이상(以上)이 요구(要求)되는 것 같다. 5. 기초대사량(基礎代謝量)의 측정결과(測定結果)는 생체중(生體重) 27.7kg의 모산양(牡山羊)을 2~3일(日) 절식(絶食)시켰을때 412Kcal 였으며 3~4일(日)에서는 240Kcal로 저하되었다. 시험기간(試驗期間)의 외기온도(外氣溫度)는 $29{\sim}34.5^{\circ}C$였으며 chamber 내(內) 온도(溫度)가 $32^{\circ}C$이상(以上)일때는 호흡(呼吸)이 panting 상태(狀態)로 되었다. 6. 재래산양(在來山羊)은 사료(飼料)의 단백질(蛋白質) 급여수준(給與水準)에 따라서 혈청내(血淸內) 단백질량(蛋白質量)은 완만하게 감소하였으나 혈청내(血淸內) 요소량(尿素量)은 현저하게 감소되는 것이 인정(認定)되었다.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2016년도 춘계학술대회 및 임시총회
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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