• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.915-921
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• 2001

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. In order to express a high level of fungal phytase in Saccharomyces cerevisiae, various expression vectors were constructed with different combinations of promoters, translation enhancers, signal peptides, and terminator. Three different promoters fused to the phytase gene (phyA) from Aspergillus niger were tested: a galactokinase (GAL1) promoter, glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter, and yeast hybrid ADH2-GPD promoter consisting of alcohol dehydrogenase II (ADH2) and a GPD promoter. The signal peptides of phytase, glucose oxidase (GO), and rice amylase 1A(RAmy1A) were included. Plus, the translation enhancers of the ${\Omega}$ sequence and UTR70 from the tobacco mosaic virus (TMV) and spinach, respectively, were also tested. Among the recombinant vectors, pGphyA06 containing the GPD promoter, the ${\Omega}$ sequence, RAmy1A, and GAL7 terminator expressed the highest phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase activity in a culture filtrate, which was estimated at 20 IU/ml. An intracellular localization of the expressed phytase was also performed by inserting an endoplasmic reticulum (ER) retention signal, KDEL sequence, into the C-terminus of the phytase within the vector pHphyA-6. It appeared that the KDEL sequence directed most of the early expression of phytase into the intracellular compartment yet more than $60\%$ of the total phytase activity was still retained within the cell even after the prolonged (>3 days) incubation of the transformant. However, the intracellular enzyme activity of the transformant without a KDEL sequence was as high as that of the extracellular one, thereby strongly suggesting that the secretion of phytase in S. cerevisiae appeared to be the rate-limiting step for the expression of a large amount of extracellular recombinant phytase, when compared with other yeasts.

• 해양환경안전학회지
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• 제26권6호
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• pp.681-688
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• 2020

해상을 통하여 운송되는 화학물질은 6,000여종에 이르며, 이러한 화학물질에는 해양을 오염시키거나 해양생물에 해로운 HNS(Hazardous and Noxious Substances)가 포함되어 있다. 해상운송 과정에서 바다로 유출된 HNS는 해상 및 해중에서 물리적, 화학적 변화를 거치게 되는데, 어떤 종류의 HNS는 해저로 침강하여 퇴적되기도 한다. 해저에 침적된 HNS는 해저생태계에 커다란 악영향을 주기 때문에 해저침적 HNS를 탐지·처리하여 회수하는 것이 바람직하다. 따라서 본 연구는 해저침적된 HNS를 회수하기 위한 기계장치를 개발할 때, 최우선으로 고려해야 하는 성능요건을 제시하였다. 해저의 오염물질을 회수하기 위하여 현재 사용되는 다양한 방식의 준설장비에 대하여 조사하고, 준설장비의 방식별 성능지표를 분석함으로써 기계장치에 대한 10가지의 성능지표를 선정하였다. 이러한 성능지표를 활용하여 해저침적 HNS 회수용 기계장치의 개발을 위한 성능요건을 제안하였다. 국내 항만의 수심을 고려하여, 해저침적 HNS 회수용 기계장치의 성능요건을 생산율(50 ~ 300 ㎥/hr), 최대운용수심(50 m), 저질 종류(대부분의 저질 형태), 고형률(10 % 이상), 수평 작업 정확도(± 10 cm), 제한 유속(3 ~ 5 knot) 등으로 제안하였다. 이러한 성능요건은 해저침적 HNS 회수용 기계장치의 개념설계와 기본설계에 활용될 것으로 기대된다.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제16권1호
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• pp.77-82
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• 2003

This investigation was conducted to study the effect of dietary formic acid (FA) and propionic acid (PA) mixture on inhibitory effect of Salmonella pullorum in layer chicks. Nine groups of one day-old layer chicks in addition to positive and negative controls, were fed with acids treated feed containing mixture of different acids concentrations, from 0.5% and 0.5% up to 1.5% and 1.5% FA and PA, respectively. Positive and negative controls were fed untreated feed. Groups except the negative control were challenged orally on day three with $10^4$ cfu/ml S. pullorum. Cloacal swabs were taken at three successive days and at 7, 14 and 21 days of challenge. After 1, 2 and 3 weeks after challenge, 4 chicks from each group were sacrificed and crop and cecal contents were examined for S. pullorum and pH. The numbers of S. pullorum positive culture from the excretion of all treated groups except groups treated with mixture of 0.5% and 0.5%, 1% and 0.5%, 0.5% and 1% FA and PA decreased significantly (p<0.05) as compared with the positive control. The mortality rates of all treated groups except the group treated with 0.5% FA and 0.5% PA were decreased significantly (p<0.05) as compared with the positive control. The treatment significantly (p<0.05) lowered the pH of the crop and cecal contents in all groups except the group treated with 0.5% FA and 0.5% PA as compared with the control. Also, the treatment significantly (p<0.05) lowered the pH of the crop and cecal contents in all groups after three weeks of treatment compared to the first and second weeks. The treatments significantly (p<0.05) lowered the frequency of S. pullorum recovery from crop and cecal contents in six groups treated with 1.5 and 0.5, 1 and 1, 1.5 and 1, 0.5 and 1.5, 1 and 1.5, 1.5% and 1.5% FA and PA respectively. These results indicate that addition of FA and PA mixture in a total concentration of 2 % or more to the diet of newly hatched infected layer chicks significantly decreases the crop and cecal colonization by S. pullorum and significantly decreases S. pullorum fecal excretion and reduced the chick mortality rate.

• Asian-Australasian Journal of Animal Sciences
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• 제12권8호
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• pp.1205-1214
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• 1999

Whole lupinus albus seeds were pressure toasted at temperatures of 100, 118 and $136^{\circ}C$ for 3, 7, 15 and 30 min to study rumen degradation and post-rumen digestion and to determine optimal heating conditions for the Dutch dairy feed industry. In sacco nylon bag and mobile bag techniques were employed for rumen and intestine incubations to determine ruminal degradation characteristics and intestinal digestion of crude protein (CP) in 4 lactation rumen cannulated and 4 lactating intestinal cannulated Dutch dairy cows fed 47% hay and 53% concentrate according to Dutch dairy requirements. Measured rumen degradation characteristics were soluble fraction (S), undegradable fraction (U), potentially degradable fraction (D), lag time (T0) and rate of degradation (Kd) of insoluble but degradable fraction. Percentage bypass feed protein (BCP), ruminal microbial protein synthesized based on available nitrogen (N_MP) and that based on available energy (E_MP), true protein supplied to the small intestine (TPSI), truly absorbed BCP (ABCP), absorbed microbial protein (AVP) in the small intestine, endogenous protein losses in the digestion (ENDP), true digested protein in the small intestine (TAP or DVE in Dutch) and degraded protein balance (PDB or OEB in Dutch) were totally evaluated using the new Dutch DVE/OEB System. Pressure toasting decreased (p<0.001) rumen degradability of CP. It reduced S (p<0.05) and Kd (p=0.06), increased D (p<0.05) and U (p<0.01) but did not alter T0 (p>0.05), thus resulting in dramatically increased BCP (p<0.001) with increasing time and temperature from 73.7 (raw) up to 182.5 g/kg DM ($136^{\circ}C/15min$). Although rumen microbial protein synthesized based on available energy (E_MP) was reduced, true protein (microbial and bypass feed protein) supplied to the small intestine (TPSI) was increased (p<0.001) from 153.1 (raw) to 247.6 g/kg DM ($136^{\circ}C/15min$). Due to digestibility of BCP in the intestine not changing (p>0.05) average 87.8%, the absorbed BCP increased (p<0.001) from 62.3 (raw) to 153.7 g/kg DM ($136^{\circ}C/15min$). Therefore DVE value of true digested protein in the small intestine was significantly increased (p<0.001) from 118.9 (raw) to 197.0 g/kg DM ($136^{\circ}C/15min$) and OEB value of degraded protein balance was significantly reduced (p<0.001) from 147.2 (raw) to 63.1 g/kg DM ($136^{\circ}C/15min$). It was concluded that pressure toasting was effective in shifting degradation of CP of lupinus albus from the rumen to small intestine without changing intestinal digestion. Further studies are required on the degradation and digestion of individual amino acids and on the damaging effects of processing on amino acids, especially the first limiting amino acids.

• Asian-Australasian Journal of Animal Sciences
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• 제10권4호
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• pp.335-356
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• 1997

Injection of bovine growth hormone (bGH) to lactating dairy cows increases milk yield and yields of milk components including fat. It is generally believed that most of the anabolic effects derived from bGH in animal tissues are primarily mediated by IGF-1. IGF-1 is a strong anabolic peptide in the plasma of animals and exerts mitogenic and metabolic effects on target cells. Contrary to most protein hormones, the majority of IGF-1 in circulation is bound to the binding proteins (IGFBPs) which are known to be responsible for modifying the biological actions of IGF-1, thus making determinations of IGF-1 actions more difficult. On the other hand, fat is a major milk component and the greatest energy source in milk. Currently, the fat content of milk is one of the major criteria used in determining milk prices. It has been known that flavor and texture of dairy products are mainly affected by milk fat and its composition. Acetyl-CoA carboxylase (ACC) is the rate limiting enzyme which catalyzes the conversion of acetyl-CoA to malonyl-CoA for fatty acid synthesis in 1ipogenic tissues of animals including bovine lactating mammary glands. In addition to the short-tenn hormonal regulation of ACC by changes in the catalytic efficiency per enzyme molecule brought about by phosphorylation and dephosphorylation of the enzyme, the long-term hormonal regulation of ACC by changes in the number of enzyme molecules plays an essential role in control of ACC and lipogenesis. Insulin, at supraphysiological concentrations, binds to IGF-1 receptors, thereby mimicking the biological effects of IGF-1. The receptors for insulin and IGF-1 share structural and functional homology. Furthermore, epidermal growth factor increased ACC activity in rat hepatocytes and adipocytes. Therefore, it can be assumed that IGF-1 mediating bGH action may increase milk fat production by stimulation ACC with phosphorylation (short term) and/or increasing amounts of the enzyme proteins (long term). Consequently, the main purpose of this paper is to give the readers not only the galactopoietic effects of bGH, but also the insight of bGH action with regard to stimulating milk fat synthesis from the whole body to the molecular levels.

• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.1041-1050
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• 2009

Equine respiratory disease is a common cause of poor performance and training interruptions. The higher incidence rate of infectious upper respiratory disease (IURD) in thoroughbred racehorses at the Seoul Race Park coincided with the frequent stabling season, shorter stabling periods, and younger ages in this study. Incidence rates were also correlated with significantly lower proportions of cells expressing MHC class II-, CD2 antigen-, $CD4^+$- or $CD8^+$-T lymphocyte-, and B lymphocyte in IURD patients compared with healthy control groups in the summer and fall and in 2-and-3-year-old groups. The data suggested that movement and new environments may have resulted in immunosuppression and inappropriate responses to respiratory pathogens in IURD patients. The IURD incidence decreased with age, perhaps by the acquisition of immunity, and study results suggested that immunologic protection was associated with IURD, particularly in young thoroughbred racehorses. Streptococci isolates were identified in 11 of 72 IURD horses, and 3 of these isolates were identified as Streptococcus. equi subsp. equi. S. equi subsp. zooepidemicus was isolated from 2 of 23 IURD horses in the spring (8.7%), 5 of 23 in the summer (21.7%), and 1 of 6 in winter (16.7%). S. equi subsp. zooepidemicus (5%) was also identified in 3 of 61 isolates from clinically normal horses. Racetracks should implement anti-IURD protective measures by assessing the capacity of equine immunologic protection at the Park and by limiting the introduction of specific respiratory pathogens (such as S. equi subsp. equi) by preventing the access of infected but subclinical horses with a specified respiratory pathogen-free certification system prior to Park entry.

• 한국콘텐츠학회논문지
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• 제16권11호
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• pp.661-674
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• 2016

최근의 스마트폰 사용 급증에 비하여 고령자의 스마트폰 사용 비율은 상대적으로 높지 않다. 노화에 의한 신체적, 인지적 능력 저하, 기존의 휴대폰에서는 볼 수 없었던 터치기반의 복잡하고 낯선 사용자 인터페이스 등의 이유로 고령자들이 자유롭게 스마트폰을 사용하는데 어려움을 겪기 때문이다. 본 연구는 스마트폰 한글 타이포그래피가 고령자의 가독성에 미치는 영향에 대해 연구하고, 고령자의 스마트폰 사용에 적합한 한글타이포그래피의 적합한 요소를 추출하고자 하였다. 65세 이상의 노인 24명을 대상으로 글자크기, 자간, 행간의 변화에 따른 판독성(Legibility) 평가결과, 서체의 종류는 나눔명조가 나눔고딕 보다 우수하였으며, 서체의 크기는 11, 12, 13pt중 13pt가 우수하였다. 반면 자간 넓이, 행간 넓이는 가독성에 영향을 미치지 않은 것으로 나타났다. 선호도 평가에서는 나눔고딕과 나눔명조는 차이를 보이지 않았다. 서체의 크기에서는 13pt, 12pt, 11pt 순으로 서체 크기가 클수록 고령자가 선호했다. 반면 자간 넓이, 행간 넓이에 따른 선호도는 차이가 없는 것으로 나타났다. 이러한 결과를 스마트폰의 한글타이포그래피에 반영한다면 고령자의 스마트폰 사용에 작게나마 도움을 줄 것으로 기대한다.

• 한국미생물·생명공학회지
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• 제45권3호
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• pp.226-235
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• 2017

Phytoene, lycopene, ${\beta}-carotene$과 같은 카로티노이드는 식품의 착색제나 영양보조제, 사료첨가제, 화장품의 원료로 사용된다. 이전 연구에서 본 연구진은 분홍색의 색소를 생산하는 새로운 해양 세균인 K. gwangalliensis를 분리 동정하였다. Phytoene desaturase (CrtI) 효소는 crtI 유전자에 암호화되어 있으며, phytoene을 lycopene으로 전환하며, 카로티노이드 합성 초기 단계에 있어서 필수적이다. CrtI는 카로티노이드 생합성 조절의 주요 효소 중 하나이며, 다양한 카로티노이드를 생합성하는 생물들의 카로티노이드 생합성 경로에 있어서 속도 조절 단계에 관련이 있다. 본 논문에서는 K. gwangalliensis로부터 lycopene 생합성을 담당하는 crtI 유전자를 클로닝 하였으며, 이 유전자는 1,584개의 염기서열을 가지며, 527개의 아미노산을 암호화하고 있다. crtI 유전자의 염기 서열을 Kocuria rhizophila와 Myxococcus xanthus를 포함한 다른 종의 염기 서열과 비교한 결과, 진화 과정에서 잘 보존되어 있음을 확인하였다. crtI 유전자를 포함하는 발현 플라스미드를 구축하여 발현시킨 결과, 이 플라스미드를 함유하는 대장균은 약 57 kDa의 재조합 단백질을 생산화였으며, 이는 phytoene desaturase의 분자량에 해당한다. lycopene의 생합성은 lycopene 생합성에 필요한 crtE, crtB 유전자를 포함한 pRScrtEB plasmid를 E. coli에 형질전환 했을 때, Escherichia coli에서 합성되는 것을 확인하였다. 이 연구의 결과는 분자 수준에서 K. gwangalliensis CrtI의 1차 구조에 대한 폭 넓은 지식 기반을 제공할 것이다.

• Korean Chemical Engineering Research
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• 제48권1호
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• pp.103-109
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• 2010

하폐수 처리과정에서 발생되는 슬러지는 주로 혐기소화에 의해 처리되며 슬러지를 감량하고 메탄을 생산할 수 있어 많이 이용되고 있다. 슬러지의 전처리는 혐기소화의 율속단계인 가수분해를 높여 처리속도를 향상시키므로 많은 연구가 진행 중이다. 본 연구에서는 열, 초음파, 열-알칼리의 전처리 기술에 따른 슬러지의 가수분해(가용화) 효과와 전처리한 슬러지를 혐기소화하여 메탄 생산량과 슬러지의 감량 효과를 비교하였다. 하수와 폐수 슬러지 가용화율은 열-알칼리 동시 처리한 경우에는 67과 70%로 가장 높았고 다음으로 초음파 처리와 열처리가 40% 이상의 비슷한 가용화율을 보였다. 혐기소화 가스의 메탄 함량은 45~70% 범위로 유지되었고 전처리한 슬러지가 control에 비해 높게 나타났다. 메탄 생산량은 열처리, 초음파 처리, 열-알칼리를 같이 처리한 경우가 control에 비해 각각 하수슬러지는 2.6, 2.7, 3.5배, 폐수 슬러지는 3.5, 4.1, 4.2배 증가하였다. 혐기소화 슬러지의 감량효과는 전처리한 슬러지가 control에 비해 5~19% 포인트 높게 나타났으며 열-알칼리 처리한 경우가 초음파와 열처리에 비해 우수한 감량 효과를 보였다. 위의 결과로부터 전처리가 메탄 생산량에서 뿐만 아니라 슬러지 처리처분 비용 절감에 있어서도 중요한 역할을 함을 확인할 수 있었고 열-알칼리 동시 처리가 가장 우수한 성능을 보였다.