• 보건의료산업학회지
• /
• 제2권1호
• /
• pp.137-154
• /
• 2008

The increase of the supply of medical service and the increase of hospitals have intensified the competition of hospitals, and the advancement towards internationalization in the opening of medical industry has triggered the infinite competition of medical profession. In addition, the high expectation of customers and quality improvement in the medical care in accordance with the improvement of overall income, and the change of active role of medical consumers according to the popularization and the improvement of rights awareness reflect the customer needs and choice in the medical service. Customers wanted to receive the kind and pleasant service under the up-to-date medical service. Therefore, as a solution, hospital coordinators were emerged for the purpose of smooth treatment and customer satisfaction by generalizing all service of hospital. Accordingly, this thesis attempted to investigate the effect of hospital coordinator education curriculum on the education satisfaction and the quality of medical service. In order to solve the purpose of this study, I, author reviewed the existing literatures, established hypothesis, and verified hypothesis by using the variety of statistics techniques such as reliability, validity, frequency analysis, and regression analysis. The verification of hypothesis is as followings: First, among education training factors of hospital coordinators, the quality of instructor significantly affects the satisfaction of hospital coordinator education training. Second, among training factors of hospital coordinator, the attitude of trainee significantly affects the training satisfaction of hospital coordinator. Third, among education training factors of hospital coordinator, education course significantly affects the training satisfaction of hospital coordinator education. As the qualities of instructor are better equipped, the satisfaction of education becomes higher. It indicates that the education method of instructors is important as an index to represent the qualities of instructor such as the appropriateness of education method, preparation, passion, visual materials, the adequacy of education procession, and specialized knowledge, and it has important effect on the satisfaction of education. In order to enhance the satisfaction of hospital coordinator education, the creation of education environment, making trainee concentrate on the education, is required by appropriately allocating programs, arousing interest in education, based on the attitude of trainee, discussion, and preliminary programs, preparation, ahead of enforcement of education. Fourth, the satisfaction of hospital coordinator education training significantly affects the reliability among the qualities of medical service. Fifth, satisfaction of hospital coordinator education training significantly affects hospitality I kindness among the qualities of medical service. If the education satisfaction of trainee is high, it is effective in the practical application such as dealing with complaints, the duty performance for the patients, and so on in offering the medical service, related to reliability and furthermore, we can find the positive change in the attitude change of medical professions related to the reliability of hospital coordinator. In addition, in the process of offering medical services such as the kind explanation on the duty, rapid response to the customers inquiry, and tidy uniform, practical effect was verified. Sixth, the education training factor of hospital coordinator significantly affects the reliability among the quality of medical service. Seventh, the education training factors of hospital coordinator significantly affect hospitality/kindness. In the education of hospital coordinator, the methods to attract the interest of trainee by emphasizing reliability should be sought and for gaining the practical effect of hospital coordinator education, the sufficient preparation and investigation on the education curriculum should be prerequisite and under this condition, intensified discussion on the instructor and education course is needed. In the design of education course, more education hours and subjects should be allocated in the part of hospitality in order to improve the practical application of hospitality. Therefore, it is meaningful in a sense that this study newly approached the components of hospital coordinator education and the need to modify the quality components of medical service in accordance with the study subjects was raised. This study also finds its meaning in that it provides basic materials for the study of future hospital coordinator education by suggesting the system development model of hospital coordinator education through preliminary study of education training. In addition, this study is meaningful in the aspect that it suggested the direction of education training by showing how the hospital coordinator education training would applied to the hospital coordinator course of the Continuing Education Center at Pusan and Kyungnam National University to some extent. Since all investigation of this study was approached from the side of hospital coordinator, the thoughts of patients who are beneficiaries of medical service, and care givers cannot be identified. Therefore, the satisfaction of patients and care givers through the experience of medical service, which is the essential prerequisite of medical service, should be importantly considered and investigated. Accordingly, The study of comparing and analyzing the views of both patients and care givers should be carried out in the future.

• 농촌의학ㆍ지역보건
• /
• 제35권3호
• /
• pp.287-300
• /
• 2010

본 연구는 지역 주민을 대상으로 하여 보건소에서 시행하는 건강증진 프로그램 참여와 관련된 요인을 파악하기 위해 시행되었다. 연구 대상은 2009년 상반기에 시행된 일개 보건소의 지역특화 건강증진 프로그램에 신청하여 기초 조사를 받은 여자 199명을 대상으로 하였으며, 이들의 건강증진 프로그램의 출석 횟수를 파악하여 참여 정도를 분류하였다. 출석 횟수에 근거하여 총 36회의 프로그램 기간 동안 전혀 참여하지 않은 비참여군과 1회 이상 출석한 참여군, 30회 미만 출석한 참여 실패군과, 30회 이상 출석한 참여 성공군등으로 분류하여 건강증진 프로그램의 참여여부와 참여 성공과 관련된 특성을 파악하였다. 건강증진 프로그램 신청자의 참여 정도를 보면, 비참여군은 28.6%이었고, 1회 이상 참여한 경우는 71.4%, 이중 30회 이상 성공적으로 참여한 경우가 28.1%였다. 보건소 시행 건강증진 프로그램참여 여부와 관련이 있었던 특성은 연령, 배우자 유무, 교육수준, 독거 여부, 월평균 소득, 프로그램장소로의 접근 방법과 동반 참여자, 평소 운동시설에의 접근성, 만성질환 이환여부와 건강생활에 대한 인지된 장애와 호감도 등이었다. 다중로지스틱 회귀분석 시행결과 건강증진 프로그램 참여 여부와는 프로그램 시행 장소로 접근 방법이 다른 교통수단을 이용한 것에 비해 도보로 접근하는 경우 교차비(95% 신뢰구간)가 4.72(1.80-12.36)로 통계적으로 유의하였고, 프로그램의 참여 성공과 관련된 특성으로는 건강생활에 대한 호감도의 교차비(95% 신뢰구간)는 1.26(1.01-1.57), 고혈압 이환 여부는 2.37(1.03-5.44), 체질량지수는 1.18(1.03-1.35)등이었다. 결론적으로 본 연구는 지역 주민이 보건소에서 제공하는 건강증진 프로그램에 참여를 결정하는 데는 프로그램이 진행되는 장소에서의 접근 방법이, 성공적으로 참여하는 데는 대상자의 건강상태와 건강생활에 대한 인식 등이 영향을 미침을 알 수 있었다. 이러한 연구 결과는 향후 보건소에서 시행하는 건강증진 프로그램을 진행하는데 있어서 참여자 관리 및 새로운 프로그램의 개설 등에 활용할 수 있을 것이고, 또한 향후 프로그램이 구성과 내용에 관한 정보를 고려하여 주민의 건강증진 프로그램 참여를 높일 수 있는 다양한 연구의 진행이 이루어져야 할 것으로 생각된다.

• Journal of Nutrition and Health
• /
• 제4권4호
• /
• pp.39-46
• /
• 1971

Iso-butyl bromide의 합성을 시작으로 D.L-1-amino-3-methylbutylphosphonic acid의 합성을, Diethyl phophite의 합성을 시작으로 D.L-1-amino-2-phenyl ethylphosphonic acid의 합성을 하였다. 합성되어진 필수 아미노산인 Isoleucine analogue phosphonic acid는 각각 0.2%, 0.4%를 식이에 첨가했고 Phenylalanine analogue phosphonic acid는 0.35%와 0.7%를 각각 첨가하였으며 Standard-1과 Standard-2군과 비교군인 Isoleucine 0.2%와 0.4% Phenylalanine 0.35%와 0.7%군으로써 총 100마리의 숫쥐로 각 7주씩 14주의 사육 실험을 하였다. 동물 사육 기간내의 뇨채취와 사육후의 각 장기의 채취로써 총 질소와 P, glycogen측정을 시도하였다. 이러한 일련의 실험결과 (1) D.L-1-amino-3-methylbuthylphosphonic acid나D.L-1-amino-2-phenylethylphosphonic acid의 유기 합성에 있어서는 다 수율이 낮고 상당히 높은 융점을 보였으며 ${\alpha}-NH_2$기가 Ninhydrin 반응에 예민했다. (2) Aminophosphonic acid의 첨가군(D.L-1-amino-3-methylbutyl phosphonic acid, D.L-1-amino-2-Phenylethylphosphonic acid)에서나 Amino acid(Isoleucine, Phenylalanine)첨가군에 있어서 모든 장기의 무게, 총체내질소 보유율, 간장내 총단백질량과 인의양, 및 뇨를 통한 인의 배설량에 각각 큰 차이를 보이지 않았다. 이로 보아서 Phenylalanine Aminophosphonic acid나 Isoleucine aminophosphonic acid가 다 보통 아미노산인 Isoleucine이나 Phenylalanine에 못지않게 체내에서 이용됨이 밝혀졌다. (3) 그러나 Phenylalanine aminophosphonic acid 첨가군이 간의 glycogen 함량에 있어서 Phenylalanine 첨가군과 Standard군과 비교해 보았을때 높았으며 이는 통계적인 유의성을 나타냈다.(<0.05) 이는 Phenylalanine Aminophosphonic Acid가 active state로써 간내에서 쉽게 이용되어서 Tricarboxylic cycle의 intermediate로 incorporate되는 경향으로 생각 할 수 있다.

• 복식
• /
• 제2권
• /
• pp.73-88
• /
• 1978

This article is an attempt to explain, at least in part, the contemporary Korean women's consciousness of Western Dreasses. As time changes, the role of clothing undergoes varisous transitions, while values and ways of life are constantly in change. It is, therefore, proper and appropriate to recognize as among the major aspects of social psychology such phenomenon as interests, understanding of clothing, the choice of a dress, and attitudes toward clothing, etc. The purpose of this study is to discover problems concerning and their clothing and their solutions, by means of a surveying approach. The method of research used is based upon questionares distributed to parents of first-year pupils in elementary schools and to female clerks working in offices, covering the period from August through October, 1976. The number of the questionares distrubuted totalled 600, and 526 were returned to the research to be utilized for analysis. The contents of the survey included such things as values concerning clothing, kinds of clothing and their practical use, the selection of clothing and the method of purchase, fashions, etc. The classification of aquisition are self-made clothing, clothing made to order and ready-made materials. It is composed of 25 items, including affirmative reasons as well as negative ones. The processing of the material returned was made by using the computer, and based upon classifications such as ages, monthly income, occupations; thus diagraming the result in percentages. The conclusion made and the improvements proposed are as follows: 1. The values of clothing were placed on the expression of the wearer's personality (32.7) and on eauty(28. 6%). The lower age group places is stress upon the expression of personality, while the higher age group stresses beauty. About 50% of wearers are contented with their clothing, their clothing, the rest of whom them indicating their dissatisfaction with what they wear. As to designs at the time of selection, about 46% indicated their preference of personal expression, 31.8% on usefulness. In selecting material, practicality is emphasized; in selecting patterns, single color is preferred. In short, personal expression and esthetic values are primary, with consideration of practicality in mind. 2. The classification of clothing according to their uses indicates the highest numbers in normal wear (home wears) and clothings to be worn outside home. As to evening dresses, (party dress) only one or two articles were checked by many, and no such article was clamed to be possessed by most. The highest ratio of wearing was shown in the case of home wear (47.3%) and clothing to be worn outside the home, which is 55.8%. The budget for one article of clothing was greatest in the case of home wear, and clothing worn outside the home. Many used both kinds of articles for the same purpose. It is desirable, therefore, that the kinds of clothing should be varied according to the purpose for which they are worn, and that clothing appropriate for that purpose should be worn. 3. The motivation for purchasing clothing was highly chosen in the item of seasonal change, which was 55.7%; Clothing deliberately made was indicated by 45.2%. In the mothods of purchasing clothing, clothing made to order and ready-made was indicated by 44.4%, which is the highest; Clothing made to order was 25.4%, and self-sewing was 1.1%, which is the lowest. (1) In the case of self-sewing, "I like it but it is very hard," was checked by 43.6%; "It is so difficult that I cannot wear such clothing" was checked by 13.3%. From these, we can conclude that the questionees are willing to make clothing by themselves, but techniques involved in sewing and at her problems involved in the skill are complicated but when those problems are eliminated there is a possibility for practice. The response checked by questionees concerning the self-sewing was, "It's economical", which is a clear indication that many questionees are positive for self-sewing. It is generally believed that ready-made clothing is cheaper, but it is not necessarily so. In consideration of the quality of clothing, self-sewing is a necessity, and it is desirable that it should be encouraged. (3) Problems involved in ready-made clothing, such as designs, skills, size (fitting) should be eliminated. When these problems are scientifically gotten rid of, it is possible that affirmative returns will be expected. Affirmative responses such as "Ready-made clothing is economical," "You can select there on the spot," are good signs that many women expect to wear ready-made clothing. It is in this sense that the prospect for ready-made clothing is brighter when much development for ready-made clothing is on the way. 4. Much concern for fashion are checked in such item of questions as "Fashionable clothing in the show window," "Clothes worn by women." The first item was checked by 50.1 %, and the second was checked by 48.6%. The reason for following fashion is "Because many people wear them," which was indicated by 30.4%. The reason for not following fashion is "It is too expensive," which was checked by 29.6%. The 26.2% of the answers indicated that "Fashionable clothing is devoid of personality," The influences of fashion over the development of fashion over the development of clothing are two-fold: Esthetic and active. It is not to be deniable that people follow fashion more or less. 1978.9>

• Asia pacific journal of information systems
• /
• 제22권2호
• /
• pp.21-38
• /
• 2012

Until recently, successful implementation of ERP systems has been a popular topic among ERP researchers, who have attempted to identify its various contributing factors. None of these efforts, however, explicitly recognize the need to identify disparities that can exist between organizational information requirements and ERP systems. Since ERP systems are in fact "packages" -that is, software programs developed by independent software vendors for sale to organizations that use them-they are designed to meet the general needs of numerous organizations, rather than the unique needs of a particular organization, as is the case with custom-developed software. By adopting standard packages, organizations can substantially reduce many of the potential implementation risks commonly associated with custom-developed software. However, it is also true that the nature of the package itself could be a risk factor as the features and functions of the ERP systems may not completely comply with a particular organization's informational requirements. In this study, based on the organizational memory mismatch perspective that was derived from organizational memory theory and cognitive dissonance theory, we define the nature of disparities, which we call "mismatches," and propose that the mismatch between organizational information requirements and ERP systems is one of the primary determinants in the successful implementation of ERP systems. Furthermore, we suggest that customization efforts as a coping strategy for mismatches can play a significant role in increasing the possibilities of success. In order to examine the contention we propose in this study, we employed a survey-based field study of ERP project team members, resulting in a total of 77 responses. The results of this study show that, as anticipated from the organizational memory mismatch perspective, the mismatch between organizational information requirements and ERP systems makes a significantly negative impact on the implementation success of ERP systems. This finding confirms our hypothesis that the more mismatch there is, the more difficult successful ERP implementation is, and thus requires more attention to be drawn to mismatch as a major failure source in ERP implementation. This study also found that as a coping strategy on mismatch, the effects of customization are significant. In other words, utilizing the appropriate customization method could lead to the implementation success of ERP systems. This is somewhat interesting because it runs counter to the argument of some literature and ERP vendors that minimized customization (or even the lack thereof) is required for successful ERP implementation. In many ERP projects, there is a tendency among ERP developers to adopt default ERP functions without any customization, adhering to the slogan of "the introduction of best practices." However, this study asserts that we cannot expect successful implementation if we don't attempt to customize ERP systems when mismatches exist. For a more detailed analysis, we identified three types of mismatches-Non-ERP, Non-Procedure, and Hybrid. Among these, only Non-ERP mismatches (a situation in which ERP systems cannot support the existing information needs that are currently fulfilled) were found to have a direct influence on the implementation of ERP systems. Neither Non-Procedure nor Hybrid mismatches were found to have significant impact in the ERP context. These findings provide meaningful insights since they could serve as the basis for discussing how the ERP implementation process should be defined and what activities should be included in the implementation process. They show that ERP developers may not want to include organizational (or business processes) changes in the implementation process, suggesting that doing so could lead to failed implementation. And in fact, this suggestion eventually turned out to be true when we found that the application of process customization led to higher possibilities of failure. From these discussions, we are convinced that Non-ERP is the only type of mismatch we need to focus on during the implementation process, implying that organizational changes must be made before, rather than during, the implementation process. Finally, this study found that among the various customization approaches, bolt-on development methods in particular seemed to have significantly positive effects. Interestingly again, this finding is not in the same line of thought as that of the vendors in the ERP industry. The vendors' recommendations are to apply as many best practices as possible, thereby resulting in the minimization of customization and utilization of bolt-on development methods. They particularly advise against changing the source code and rather recommend employing, when necessary, the method of programming additional software code using the computer language of the vendor. As previously stated, however, our study found active customization, especially bolt-on development methods, to have positive effects on ERP, and found source code changes in particular to have the most significant effects. Moreover, our study found programming additional software to be ineffective, suggesting there is much difference between ERP developers and vendors in viewpoints and strategies toward ERP customization. In summary, mismatches are inherent in the ERP implementation context and play an important role in determining its success. Considering the significance of mismatches, this study proposes a new model for successful ERP implementation, developed from the organizational memory mismatch perspective, and provides many insights by empirically confirming the model's usefulness.

• Journal of Chest Surgery
• /
• 제42권1호
• /
• pp.22-27
• /
• 2009

배경: 관상동맥우회술은 관상동맥질환의 대표적인 치료법으로 서구에서는 장기 성적에 관하여 많은 보고들이 있으나, 우리나라에서는 매우 드물다. 따라서, 본원에서의 장기 임상성적에 관하여 알아보고자 하였다. 대상 및 방법: 1984년 2월 본원에서 처음으로 관상동맥 우회로술을 시행한 이후 2006년 12월 31일까지 시행하였던 342명의 환자를 대상으로 하였으며, 의무기록을 토대로 후향적으로 분석하였다. 총 286명(83.6%)에서 추적관찰이 가능하였으며, 평균추적관찰기간은 $75.7{\pm}46.1$개월이었다. 결과: 조기사망은 5.6%였으며, 만기사망은 23.1%였다. 1년 생존율은 91.5%였으며, 5년 생존율은 82.1%, 10년 생존율은 60.7%, 15년 생존율은 50.0%였다. 60세 이상의 고령의 환자(p=0.002), 당뇨(p=0.000), 고혈압(p=0.002), 다중혈관질환 환자(p=0.006), 그리고, 좌심실 기능부전 환자들(p=0.015) 환자들에서 생존율이 유의하게 낮았으며, 성별은 장기생존에 차이가 없는 것으로 나타났다. 다변량 분석에서는 당뇨(p=0.001), 나이(p=0.005), 좌내흉동맥을 사용하지 않는 경우(p=0.037)가 유의한 위험인자로 나타났다. 결론: 관상동맥우회술은 관상동맥질환 환자에게 있어서 효과적인 치료방법이라고 생각되며, 장기 생존을 위해서는 내흉동맥의 적극적인 사용과 당뇨환자에서 술후 혈당조절과 같은 적절한 약물치료와 외래추적관찰이 필요할 것으로 생각된다.

• 농약과학회지
• /
• 제20권3호
• /
• pp.271-279
• /
• 2016

전신복장법(whole body dosimetry, WBD)을 이용한 사과 과수원 농약 살포자의 살포농약 조제 및 살포 과정 중 imidacloprid 수화제 사용에 의한 노출 특성 및 위해성을 평가하기 위하여 농약 살포 전 방제복, 장갑, 마스크, 호흡 노출량 측정 장치를 착용한 후 speed sprayer를 이용하여 acephate+imidacloprid 25(20+5)% 1,000배 희석액을 3,000 L/ha/살포자의 비율로 청주 인근의 10개 사과 과수원에 살포하였다. 노출시험은 조제자와 살포자에 대하여 각각 나누어 수행하였다. 시험농약은 HPLC-DAD를 이용하여 분석하였으며, 부위별 평균 회수율은 81.5-108.6%, 평균 포장회수율은 73.8-86.7%로서 모두 적합하였다. 조제자 및 살포자의 노출량은 imidacloprid 총량 대비 0.0014-0.0279%이었으며, 조제자와 살포자 모두 장갑의 노출량이 가장 높았다. 또한 농약 살포자의 농약 사용에 대한 위해성을 측정하기 위한 안전한계(margin of safety, MOS)는 조제자와 살포자 각각 97-355, 46-196으로서 speed sprayer를 이용한 imidacloprid 살포 시 사과 과수원 농약 살포자에 대한 위해도는 매우 낮았다.

• The Korean Journal of Physiology
• /
• 제9권1호
• /
• pp.13-22
• /
• 1975

From the study of movements of $Ca^{++}$ in frog cardiac muscle, Niedergerke (1963) postulated that $Ca^{++}$ necessary for the cardiac contraction is stored in a specific pool. Langer et al (1967) and DeCaro (1967) also found a close relationship between the change of $Ca^{++}$ flux kinetics and the change of contractile force. According to the studies of several investigators, Ca II (Bailey and Dressel 1968) or phase I and II (Langer 1965, Langer et al 1967, 1971) in the $Ca^{++}$ washout curve was associated with cardiac contractility. This investigation was aimed to elucidate the anatomical region of the contractile active $Ca^{++}$ pool. At the same time, it was assumed in this study that $Ca^{++}$ in the sarcoplasmic reticulumn represents one of the major intracellular $Ca^{++}$ pool and cardiac contractility was also dependent on the intracellular $Ca^{++}$ concentration. Consequently, this experiment was performed at different temperatures to activate to activate inhibit the deactivating process of activated $Ca^{++}$ in the intracellular space to see if changes in the contractility decay curve existed at different temperatures. The isolated hearts of rabbits and turtles (Amyda maackii) were attached to the perfusion apparatus according to the method employed by Bailey and Dressel (1968). The isolated hearts were initally perfused with a full Ringer solution containing 2 mg/ml of inulin for 1 hr, and then $Ca^{++}$ and inulin-free Ringer solution was perfused while the isometric tension was recorded and a serial sample of perfusion fluid dripping from the cardiac apex was collected for 10 sec throughout experimental period. The above procedure was performed at $23^{\circ}C$, $30^{\circ}C$ and $38^{\circ}C$ on the rabbit heart and $10{\sim}13^{\circ}C$, $10^{\circ}C$, $25^{\circ}C$, $30^{\circ}C$ and $35^{\circ}C$ on the turtle heart. After determination of $Ca^{++}$ and inulin concentration of the samples, the $Ca^{++}$, inulin washout curve and the contractile tensin decay curve were analysed according to the method of Riggs (1963). The results were summarized as follows; 1. In the rabbit heart, there are 2 inulin compartments, 3 $Ca^{++}$ compartments and sing1e exponential decay of contractile tension. In the turtle heart, there are $1{\sim}2$ inulin compartments, $1{\sim}2$ $Ca^{++}$ compartments and $1{\sim}2$ phases of contractile tension decay. The fact that the inulin space was divided into 3 compartments in the washout curve in these hearts indicates the presence of heterogeneity in cardiac perfusion, i.e., overfused and underperfused area. 2. Ca I a9d Ca II in these hearts were found to have $Ca^{++}$ in the ECF compartments because their half times in the washout curves were far smaller than those of the inulin washout curves in the rabbit heart and similar to those of the inulin washout curves in the turtle heart. Ca III in the rabbit heart may have originated from the intracellular $Ca^{++}$ store. But no Ca III in the turtle heart was found. This may be due to the fact that the iutracellular $Ca^{++}$ pool in the turtle heart was too small to detect using this experimental procedure since sarcoplasmic reticulumn in the turtle heart is poorly developed. 3. In the rabbit heart, there were no chages in the half time of Ca I, Ca II, inulin I and inulin II at different temperatures, but the half time of Ca III was significantly prolonged at lower temperatures, and the half time of the contractile tension decay tended to be prolonged at lower temperatures but this was not significant. In the turtle heart, there were no changes in the half time of Ca I, Ca II, inulin 1, inulin II and phase I of the contractile tension decay at different temperatures, but the half time of phase II of the contractile tension decay was significantly prolonged at lower temperatures. This finding indicates that intracellu!ar $Ca^{++}$ in these hearts was also responsible particulary for maintaining the cardiac contractility at the lower temperatures. 4. The half times of contractile tension decay were shorter than those of Ca II in the $Ca^{++}$ washout curves in both animal hearts. According to the above results it was shown that $Ca^{++}$ in ECF is primarily and $Ca^{++}$ in the intracellular space is partially associated with the cardic contractility.

• KSBB Journal
• /
• 제25권2호
• /
• pp.155-166
• /
• 2010

Inonotus obliquus 균사체의 액상배양을 통해 항당뇨 및 면역증강 효능의 세포벽다당체 (IPS) 생산 공정을 개발하기 위한 첫 시도로서, 고생산성 균주를 개발하는 연구를 수행하였다. Inonotus obliquus는 균사체로 성장할 때 포자를 형성하지 못하므로 단일 세포주를 얻기가 힘든 것으로 관찰되었다. 따라서 Inonotus obliquus 균사체로부터 대량의 원형질체 형성 및 재생에 의한 단일 콜로니 획득 방법을 개발함으로써 생산균주를 신속하게 개량하고자 하였다. 개량된 filtration 방법을 적용해서 원형질체를 회수한 결과, 기존의 trapping 방법에 의해 회수한 수보다 약 5배 증가한 $2.3{\times}10^6$ protoplasts/mL의 원형질체를 회수할 수 있었으며, 원형질체 재생률 또한 $10^{-2}{\sim}10^{-3}$로서 비교적 높게 나타났다. 한편 Inonotus obliquus 균사체의 경우 IPS의 함량이 세포 무게 당 거의 일정한 양을 함유하고 있는 것으로 확인되었으므로, IPS의 생산성을 증가시키기 위해서는 최종 액상 생산배양에서의 균체량 증가가 가장 중요한 것으로 판단되었다. IPS 고생산성의 균주를 개발하기 위해, 이미 선별된 고생산성 균주들의 균사체를 다양한 조건으로 UV 처리한 후, 생존한 원형질체로부터 고생산성의 변이주들을 지속적으로 선별한 결과, 16~18 g DCW/L 범의의 높은 균체 생산성을 보이는 우량균주들을 다수 선별할 수 있었다. 특히 고체 성장배지에서 빠른 성장속도를 보여주는 대부분의 균주들이 최종 액상 생산배양에서도 고생산성 및 고안정성을 보여주는 것으로 나타났다. 이 결과로부터 일련의 균주 개발 공정의 초기 단계에 속하는 고체 성장배양에서 성장속도가 낮은 균주들을 미리 제외시킬 경우, 균주선별의 효율성이 매우 높아짐을 확인할 수 있었다. 최종적으로 97%의 치사율을 보이는 UV 변이처리 조건에서 균사체 생산성이 약 22 g/L에 이르며, 생산 안정성도 높은 우량 균주 (OBLQ756-15-5)를 획득할 수 있었는데, 이 균주를 이용하여 현재 IPS 대량 생산을 위한 pilot-scale 발효조 배양공정을 개발 중이다.

• Korean Chemical Engineering Research
• /
• 제47권2호
• /
• pp.220-229
• /
• 2009

본 연구에서는 Phellinus linteus 균사체의 액상배양을 통한 면역증강 생리활성 효능의 단백다당체 생산공정을 개발하기 위한 시도로서, 우선 생산균체의 원형질체 형성을 통한 고생산성 균주를 개발하고자 하였으며, 발효기 액상배양 시 최적 배양형태의 유도를 통해 균사체와 단백다당체의 생산성을 극대화하고자 하였다. 본 연구실에서 채취한 생산 균주를 ITS rDNA sequencing 방법과 blast search 방법에 의해 조사한 결과 다양한 Phellinus linteus 종들과 99.67% 이상의 유사성 확인되어, 이 균주를 Phellinus linteus라고 최종적으로 동정할 수 있었다. 이 동정된 균주로부터 균주 개량을 시도하기 위해 Phellinus linteus 균사체로부터 대량의 원형질체 형성 및 재생에 의한 단일 콜로니 획득 방법을 개발함으로써 균주를 신속하게 개량할 수 있었다. Sorbitol을 이용한 banding filtration 방법을 이용하여 원형질체를 회수한 결과 $10^5{\sim}10^6\;protoplasts/ml$를 얻을 수 있었으며, 원형질체 재생률은 $10^{-2}{\sim}10^{-3}$로 나타났다. 균주개량을 위해 원형질체 재생배지와 고체배양배지에서 고성장성 및 고안정성을 보이는 균주들을 지속적으로 대량 선별하여, 액상 생산배양을 수행하였다. 그 결과 균사체량은 13~15 g/L로 대부분 비슷하게 자랐으며, 조단백다당체의 함량 또한 5.8~6.4%로 거의 비슷하게 분포하는 것으로 나타났는데, 이로부터 고체배양배지에서 빠른 성장속도를 보여주는 균주들이 대부분 액상 생산배양에서도 고생산성 및 고안정성을 보여주는 것을 확인할 수 있었다. 한편 Phellinus linteus 균사체의 경우 조단백다당체의 함량이 세포 무게당 거의 일정한 양을 함유하고 있는 것으로 확인되었으므로, 조단백다당체의 생산성을 증가시키기 위해서는 최종 생산배양에서의 균체량 증가가 가장 중요한 것으로 판단되어, 균사형성 고등균류의 균사체 배양 시 균체량 증가에 가장 중요한 요인 중의 하나인 생산균주의 배양형태적 특성에 대해 집중적으로 조사하였다. 균주개량 실험을 통해 고생산성 균주로 최종 결정된 AR147 균주를 이용해서 다양한 배양조건에서 발효조 배양을 수행한 결과, 최종 생산발효조로의 접종원이 고농도의 균사모양인 경우에 생산균주의 배양형태가 매우 작은 compact한 펠렛 모양(대부분 직경 0.5 mm 이하)을 유지하는, 이상적인 균사체 액상배양 공정이 이루어지는 것으로 확인되었다. 즉 생산 발효조배양에서 직경 0.5 mm 이하의 compact한 펠렛 모양의 배양형태가 유도되었을 경우, lag phase 시간의 획기적 감소와 1.5배 이상의 높은 세포비성장속도로 인해, 최종 균사체생산성이 다른 배양형태를 유도한 경우에 비해 약 3.3배 더 높은 주목할 만한 배양결과를 얻을 수 있었다. 이로부터 균사 형성 Phellinus linteus의 산업용 발효조 배양 시, 각 배양단계에서의 생산균체의 배양형태가 최종 균체생산성, 궁극적으로는 최종 단백다당체의 생산성에 심각한 영향을 미친다는 것을 알 수 있었다.