• Applied Biological Chemistry
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• v.19 no.3
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• pp.162-171
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• 1976

The determination of rate limiting step in the metabolic process is of great importance to understand the metabolic properties. In this paper the authors propose a modified enzymatic method instead of thermodynamic method. This method is based on the assumption that the over-all rate increment would be larger than by any other steps to which the individual enzyme are added respectively, provided that the enzyme participated in the rate limiting step is added to the reactions composed of n steps of metabolic process with which n kind of enymes are concerned. The present paper deals with analysis and discussion about some factors having influence on the proposed process, mainly about the metabolic process constituted with homogeneous steps. The results show that the determination of rate limiting step by a modified enzymatic method is feasible, provided that some restrictions are added in any type of mechanisms.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.219-233
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• 1999

Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.208-214
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• 1994

We screened many species from a wide variety of bacterial genera for a new type II restriction endonuclease. The purification and characterization of SolI from a soil isolate, Streptoverticillium olivoverticillatum are described here. The enzyme turned out to be an isoschizomer of BamHI. It recognized the hexanucleotide sequence of 5'-G$\downarrow$GATCC-3' and cleaved as in dicated by the arrow, generating a 4 base 5' extension. Unlike its isoschizomer, BamHI, the activity was sensitive to dam methylation within the recognition sequence. Following ammonium sulfate fractionation of the crude extract, heparin-agarose and Affi-gel Blue column chromatography were employed to purify the enzyme. SolI required at least 0.2 mM of $MgCl_2$ for the cleavage to occur. The enzyme exhibited its maximal activity in the absence of NaCl, but was inhibited completely in the presence of 120 mM NaCl. The pH and temperature optima for activity were pH 8.6 and $40^{\circ}C$, respectively. The molecular weight of SolI was estimated to be 43,000 Da by Superose-12 gel filtraion chromatography.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.119-126
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• 1997

Acanthnmoebn sp. YM-4 is simitar to A. culbertsoni based upon morphological characteristics of trophozoites and cysts. However, based on other characteristics, pathogenicity to mice, in uitro cytotoxicity and isoenzyme patterns, Acanthomoebo sp. YM- 4 was quite different from A. culbertsoni. Restriction fragment length polymorphism (RFLP) analysis of mtDNA is useful in the classification of members belonging to the genus Acanthcmoebn. Therefore, in this study, RFLP analysis of Acnnthcmoeba mtDNAs was accomplished using five restriction enzymes: Hnelll, Hinull, Clcl, Pudl and ScE. Each restriction enzyme produced approximately 3-15 fragments (range: from 0:6 kip to 34.4 kbp) . The mtDNA genome size, calculated by the summation of restriction fragments, averaged 46.4 kbp in Acnnthamoeba sp. YM-4,48.3 kbp in A. culbertsoni and 48.8 kbp in A. polyphaic, respectively. Digested mtDNA fragments of Accnthcmoeba sp. YM-4 contained nine and seven same size fragments, respectively, from a total of 67 and 69 fragments observed in A. culbertsoni and A. polyphcgn. An estimate of the genetic divergence was 10.1% between Acanthamoebc sp. YM-4 and A. culbertsoni, and 9.9% between Acanthamoebn sp. YM-4 and A. polyphcga.

• Korean Journal of Food Science and Technology
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• v.51 no.4
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• pp.348-355
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• 2019

This study investigated the impact of cellulase treatment on the extraction yield of potato starch (PS), and compared the physicochemical properties of PS by conventional (CSE) and enzymatic (ESE) starch extraction. In ESE, the PS extraction yield was predominantly influenced by reaction temperature, time and their interaction, compared to the cellulase concentration. When potatoes were treated for 8 h at $40^{\circ}C$ with 1.5% cellulase, the PS extraction yield was about 3.4-fold higher than that by CSE. Compared to CSE-PS, ESE-PS showed lower total starch contents and higher amylose contents, resulting in lower swelling factors and distorted pasting viscosity profiles accompanied by absence of peak and breakdown viscosities. However, ESE did not affect the gelatinization characteristics of PS. Overall results suggested that ESE can provide the highest yield of PS, and ESE-PS can be a potential starch source for extending the utilization of PS in food industries.

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.262-266
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• 2000

The polymerase chain reaction was used to selectively detect sequences within the fimbrial antigen of Salmonella enteritidis. Sterile milk was artificially inoculated with known amount of S. enteritidis and then DNA was extracted with guanidine thiocyanate/phenol/chloroform, followed by PCR. A detection limit of as few as 100 colony forming unit (cfu) per 0.5 ml milk was obtained with this method. For the whole procedure, it took only 5 h. A semi-quantitative polymerase chain reaction assay which allows an estimation of colony forming unit of S. enteritidis was developed. Known amount of standard plasmid pGem-4Z-Sef B(-) containing cloned S. enteritidis fimbrial antigen gene was co-amplified with Salmonella genomic DNA isolated from artificially inoculated milk. The same set of primers were used for the amplification and the products were cleaved with Bam HI. The concentration of the target DNA could be estimated by comparing the intensity of the two bands after electrophoresis. The PCR-based protocol described in this paper provides a rapid, simple, and sensitive method for detecting S. enteritidis in milk.

• Microbiology and Biotechnology Letters
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• v.16 no.5
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• pp.363-368
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• 1988

Molecular cloning of gene for $\alpha$-acylamino-$\beta$-lactam acylhydrolase (ALAH) III from Acetobacter turbidans has been attempted by immunochemical detection method, in which polyclonal antibody from mouse Balb/c against this enzyme was employed as a probe. As a cloning vector, λ gtll was chosen for this purpose. Two positive clones has been selected from genomic libraries of A. turbidans, which had somewhat different binding affinities on anti-ALAH III umm and anti-$\beta$-galactosidase. By restriction analysis, both clones has been turned out to lose one of EeoRI sites. From these results, it concluded that deletion of DNA between lacZ gene and inserted DNA has occurred during replication of these clones in host cells.

• Applied Biological Chemistry
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• v.37 no.1
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• pp.56-63
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• 1994

Oligonucleotides for a conserved region of the coat protein gene of cucumber mosaic virus (CMV) and a hammerhead structure ribozyme against CMV RNA were synthesized using a DNA synthesizer. Both strands of oligonucleotides were annealed and restricted with BamHI/SacI, then cloned into a plasmid pBS SK (+). The cloned CMV substrate and ribozyme were sequenced to verify correct constructions. In vitro transcriptions were carried out by using T7 RNA polymerase with BssHII or SspI digests of $1\;{\mu}g$ of substrate and ribozyme clones. The size of substrate RNA was 176 nucleotides (nt) containing 50 nt of CMV RNA sequence, 6 nt of XbaI restriction site and 120 nt of vector-derived sequence in the case of BssHII digest. The size of ribozyme RNA was 164 nt containing 40 nt of ribozyme RNA sequence and same sequences of substrate. Substrate RNA was efficiently cleaved into two fragments (96 nt and 80 nt) by ribozyme RNA. This endonucleolytic cleavage occurred more efficiently at $55^{\circ}C$ than $37^{\circ}C$. SspI digest-derived substrate RNA (2234 nt) was also cleaved into two fragments by the same ribozyme. SspI digest-derived ribozyme RNA (2222 nt) cleaved the above substrate to two fragments. In vitro-tested ribozyme construct is being cloned into a plant transformation vector to develop virus-resistant plants.

• Journal of environmental and Sanitary engineering
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• v.14 no.1
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• pp.103-115
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• 1999

생물학적 처리공정내 미생물은 물리화학적, 생태학적으로 처리공정의 운전조건과 유입하·폐수 특성에 따라 존재량 및 반응조내 존재하는 우점종이 달라진다. 특히,폐수의 생물학적 처리시 유입수내 존재하는 독성 및 저해물질은 생물반응조내 존재하는 미생물의 양을 감소시키거나 미생물의 종류를 단순화시키거나 환경조건 및 폐수 특성변화에 능동적으로 대처할 수 있는 능력을 감소시킨다. 따라서 이러한 문제점을 극복하기 위한 방안으로 생물능력향상(bio-augmentation)이 필요하며, 여러 가지 방법으로 생물능력향상을 이룰 수 있다. 본 연구에서는 생물능력향상을 위하여 기존의 효소활성이 강한 미생물군을 투입함으로써 처리효율향상에 미치는 영향 및 제한성을 고찰하였다. 처리효율 향상을 위해서는 생존성, 폐수에 대한 순응기간 및 미생물의 체류가 매우 중요하였으며, 특히 누입된 미생물군이 처리효율을 향상시키기 위해서는 일정기간 이상의 순응기간이 필요하였다. 한편, 토착미생물군인 UB-1의 경우 기존 생물능력향상 미생물군과 거의 동일한 처리효율을 나타내고 있으며, 사전에 순양기간을 거친 UB-2의 경우도 기존 생물능력향상 미생물군과 비슷한 처리효율을 보였다. 따라서 본 결과로 볼 때 생물능력향상 미생물군 투입에 의한 처리효율의 향상은 크지 않으며, 하·폐수의 생물학적 처리공정에 적요하는데는 다소 제한성이 있은 것으로 판단되었다.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.3 no.1
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• pp.22-27
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• 2003

고페닐알라닌혈증의 경우 모든 환자들이 저페닐알라닌 식이를 엄격히 지키는 것은 아니기 때문에 기대만큼의 치료효과를 거두지 못하는 경우가 종종 있다. 모든 페닐알라닌 수산화 효소 결핍에 의한 페닐케톤뇨증 환자에서 테트라하이드로바이오프테린의 복용이 효과를 거두는 것은 아니나 식이조절과 비교해 볼 때 이는 식이 제한에 대한 어려움이 없고 복용이 간편하므로 치료효과가 있다면 일부 환자에서라 하더라도 페닐케톤뇨증 환자의 치료에 새로운 방향을 제시하는데 충분한 의미가 있다고 본다. 이러한 시점에서 저자들은 고전적 페닐케톤뇨증으로 진단되었던 6명의 환아에서 식이제한없이, 또는 식이제한을 완화한 상태에서 테트라하이드로바이오프테린을 복용함으로써 치료 효과를 거둔 6례에 대해서 문헌 고찰과 함께 보고하는 바이다.