WO2005097972A1 - 乳酸菌培地 - Google Patents
乳酸菌培地 Download PDFInfo
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- WO2005097972A1 WO2005097972A1 PCT/JP2005/004812 JP2005004812W WO2005097972A1 WO 2005097972 A1 WO2005097972 A1 WO 2005097972A1 JP 2005004812 W JP2005004812 W JP 2005004812W WO 2005097972 A1 WO2005097972 A1 WO 2005097972A1
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- Prior art keywords
- medium
- yeast extract
- whey
- food
- lactic acid
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- XAZKFISIRYLAEE-UHFFFAOYSA-N CC1CC(C)CC1 Chemical compound CC1CC(C)CC1 XAZKFISIRYLAEE-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
Definitions
- the present invention relates to a lactic acid bacteria culture medium containing whey produced as a by-product of cheese manufacture or casein manufacture. More specifically, the present invention relates to a lactic acid bacteria culture medium characterized by containing whey enriched with a nitrogen source.
- Whey produced as a by-product of cheese manufacture and casein manufacture is a highly nutritious substance containing lactose, water-soluble proteins, minerals, organic acids, water-soluble vitamins, and the like. Wheichi is also used as a concentrated whey powder for whey health foods and beauty products used as confectionery materials and lactose ingredients as whey powder. However, only a part of the whey produced in large quantities is in trouble.
- acidophilus group (6 species of L. acidophilus, L. gassen, L. amyiovorus, L. cnspatus, L. gallinarum, L. johnsonii) derived from human intestine
- acidophilus group 6 species of L. acidophilus, L. gassen, L. amyiovorus, L. cnspatus, L. gallinarum, L. johnsonii
- the acidophilus group generally grows slowly in milk, for example, when it is used for products such as yogurt, it can be grown in a medium other than the product such as yogurt and the bacteria added to the product. It is often done.
- MRS medium for culturing these lactic acid bacteria.
- This medium is very expensive (more than 30,000 yen per kg) and is difficult to use industrially. Therefore, an inexpensive and excellent medium that can replace MRS medium is desired.
- Wheichi is cheap with high nutritional value. In addition, it can be used as a safe food ingredient because it is a component that provides milk power. Therefore, if a lactic acid bacteria culture medium that can be converted into an MRS culture medium can be obtained by effectively using the whey that has been discarded, an inexpensive and industrially available medium can be obtained. Can be obtained.
- whey as a culture medium for microorganisms such as lactic acid bacteria
- examples of the use of whey as a culture medium for microorganisms such as lactic acid bacteria include a culture apparatus for lactic acid bacteria using a medium in which yeast extract is added to whey (Patent Document 1), sweet whey and yeast extract, and casein hydrolysis.
- Patent Document 1 a culture apparatus for lactic acid bacteria using a medium in which yeast extract is added to whey
- sweet whey and yeast extract a medium mixed with food, etc.
- casein hydrolysis Using a medium mixed with food, etc., the growth of urease-negative bacteria can suppress the generation of odor from animal excrement (Patent Document 2). Is not described, and a lactic acid bacteria culture medium using whey as a raw material which can be changed to an MRS medium has not been obtained.
- Patent Document 1 JP-A-6-253816
- Patent Document 2 Japanese Translation of Heisei 10-507358
- the problem to be solved by the present invention relates to the provision of an inexpensive and safe lactic acid bacteria medium. More specifically, the present invention relates to providing a lactic acid bacterium medium that can be converted into an MRS medium using whey as a raw material.
- the present inventors have prepared whey as a nutrient source that has the original strength in order to effectively use whey as a raw material for lactic acid bacteria culture medium. It was found that a medium capable of efficiently cultivating lactic acid bacteria can be obtained by strengthening the source. Furthermore, it has been found that by replacing the additive components with approved food additives, it is possible to obtain a medium that has only the ingredients safe for the human body.
- the low-cost and safe medium obtained here enables not only large-scale cultivation of lactic acid bacteria, but also the development and production of foods, pharmaceuticals, etc., using lactic acid bacteria cultured on this medium. An industrially valuable medium can be obtained.
- the medium of the present invention is characterized by strengthening the nitrogen source of the medium in addition to the nutrient source of whey.
- the whey with enhanced nitrogen source in the present invention means both enhancing the nitrogen source of whey by enzymatic decomposition of whey itself and adding a substance that becomes a nitrogen source.
- yeast extract and / or enzymatic degradation products of whey and casein It is characterized by strengthening the nitrogen source of a culture medium by containing.
- an inexpensive and safe medium can be obtained by using only food additives approved for addition.
- the present invention relates to a medium according to any one of (1) and (13) below and a culture method using the medium.
- a medium comprising whey enriched with a nitrogen source.
- Oleic acid compound power Tween80, sodium oleate, sucrose oleate ester, glycerinoleate (1)
- Tween80 sodium oleate
- sucrose oleate ester sucrose oleate ester
- glycerinoleate One of the media according to (7).
- the invention's effect [0010]
- the medium of the present invention is cultivated by this medium, which makes it possible to culture a large amount of lactic acid bacteria inexpensively and safely by using whey and approved food additives as raw materials. It is possible to develop and manufacture foods and medicines using lactic acid bacteria that are highly industrially and economically effective.
- FIG. 1 is a graph showing the growth of a test bacterium in an enzyme-degraded whey medium by a change in turbidity. (Test Example 1)
- FIG. 2 A graph showing the growth of the test bacteria in the enzyme-degraded whey medium according to the difference in the enzyme reaction time, by the change in turbidity.
- FIG. 3 is a graph showing the growth of the test bacteria in the casein enzyme-decomposed product-added medium according to the change in turbidity depending on the amount of casein enzyme-decomposed product added. (Test Example 2)
- FIG. 4 A graph showing the viability of various test bacteria by changes in turbidity.
- FIG. 5 is a graph showing the growth of test bacteria in whey medium supplemented with various oleic acid compounds by changes in turbidity. (Test Example 3)
- FIG. 6 is a graph showing the growth of test bacteria in whey medium supplemented with various oleic acid compounds by changes in turbidity. (Test Example 3)
- FIG. 7 is a graph showing the growth of test bacteria in a whey medium supplemented with a yeast extract for medium or food, as a result of changes in turbidity. (Test Example 3)
- FIG. 8 is a graph showing the growth of the test bacteria in a whey medium to which a yeast extract for medium or food is added, by the change in turbidity. (Test Example 3)
- FIG. 9 is a graph showing the growth of the test bacteria in a whey medium supplemented with a yeast extract for medium or food, as a change in turbidity. (Test Example 3)
- FIG. 10 is a graph showing the viability of various test bacteria in whey culture medium according to the change in turbidity depending on the difference in yeast extract and the amount added. (Test Example 3)
- FIG. 11 is a graph showing the viability of the test bacteria in various media in terms of dry cell weight. (Test Example 4)
- FIG. 12 is a graph showing the growth of the test bacteria when cultivated in a jar mentor and an incubator as a result of changes in turbidity. (Test Example 4) BEST MODE FOR CARRYING OUT THE INVENTION
- the preparation of the medium of the present invention is characterized in that the nitrogen source in the medium is increased by a yeast extract and / or a proteolytic enzyme in addition to the nutrient source that whey also has vitality.
- protein enzyme degradation products include enzymatic degradation whey obtained by enzymatic degradation of whey to increase the content of whey-derived protein degradation products, and casein enzyme degradation products obtained by enzymatic degradation of casein. It is done.
- the concentration of the yeast extract contained in the medium is not particularly limited as long as it is an amount effective for the lactic acid bacteria to be cultured, but in order to obtain a high growth promoting effect that is preferably 0.3% or more. 0.6% or more, more preferably 0.9% or more.
- koji is approved as a food additive and can be used as a miscellaneous protease.
- Protease P can be used.
- yeast extract is sold for food use, any deviation can be used.
- yeast extract Kyowa C yeast extract Kyowa H, yeast extract Kyowa L, yeast extract Kyowa W, yeast Yeast extract such as extract Takeda HR, yeast extract Takeda Sl ⁇ -W and yeast extract JT-21A can be used.
- Tween80 As an oleic acid compound, Tween80 has been approved as a food additive !, so! /, And so are the approved food additives such as sodium oleate, sucrose oleate, decaglycerin monooleate and others It is more preferable to use glycerol ester of oleic acid (glycerolinate) as an alternative to Tween80! By using these components, an inexpensive and safe medium can be provided.
- glycerol ester of oleic acid glycerolinate
- the medium of the present invention may further contain MgSO 4, MnSO 4, K ⁇ , ( ⁇ ) C ⁇ ⁇ ⁇ as necessary.
- NaC ⁇ ⁇ , etc. can also be exempt from calories.
- a method of removing the protein by ultrafiltration can be used in addition to coagulation and removal of the protein by heating. Thereby, a clear culture medium can be prepared.
- composition and preparation method of the MRS medium used for comparison in the lactic acid bacteria medium of the present invention Refer to the composition and preparation method of the MRS medium used for comparison in the lactic acid bacteria medium of the present invention.
- the following is an example.
- MRSBROTH MRS powder
- Table 1 shows the composition of the MRS medium.
- lactic acid bacteria culture medium of the present invention and a method for culturing lactic acid bacteria using this culture medium are shown in the following examples.
- 125 g of cheese whey powder was dissolved in 375 ml of distilled water to prepare a 25% whey solution. This is sterilized by heating in an autoclave (110 ° C, 15 min), cooled and stirred, and then centrifuged (8000 rpm, 5 ° C, 15 min) using a cooling high-speed centrifuge, and the supernatant is made up of two sheets of filter paper. And suction filtered to remove the heat-denatured protein.
- Distilled water was added to the deproteinized whey solution to dilute it 3.5 times (solid content approx. 7%), pH was adjusted to 6.5 with lN-NaOH reagent, and used as the basic whey medium.
- Glucose is D (+)-Glucose (Dextrose, Anhydrous: Wako Pure Chemical Industries, Ltd.) c, Yeast extract
- Yeast Extract (Batato) for culture media Yeast Ek Kyowa C, Yeast Extract Kyowa H, Yeast Extract Kyowa L, Yeast Extract Kyowa W (all Kyowa Hakko Kogyo Co., Ltd.)
- Yeast Extract Takeda HR Yeast Extract Takeda Sl ⁇ -W (both Takeda Kirin Foods Co., Ltd.) or Yeast Extract JT-21A JT Foods Co., Ltd.) were used.
- the oleic acid compound is Tween 80 (Difco) and the approved food additive sodium oleate (Kanto igaku), lyoto sugar ester 0-1570 (food additive Sugar oleate (Mitsubishi Ishigaku Hoos, Inc.) or decaglycerin monooleate (Riken Vitamin Co., Ltd.) was used.
- glucose was added to the basic whey medium prepared in Reference Example 2.
- Casein enzyme degradation product and yeast extract were added, and as the oleic acid compound, either sodium oleate, lyoto sugar ester 0-1570 or decaglycerin monooleate was added to prepare a medium. .
- oleic acid compound-free medium and Tween 80-added medium were prepared. The composition of the medium is shown in Table 4.
- G glucose
- C casein enzyme digest
- YEB Yeast Extract (Batato) for medium
- YE yeast extract for food addition
- 0 oleic acid compound.
- T80 Tween80
- SO sucrose oleate
- ⁇ sodium oleate
- 0 ⁇ 1 de force glycerin monooleate
- W yeast extract Kyowa W.
- Yeast Extract substitute medium for medium is supplemented with glucose, casein degradation product, and Tween 80 in basic whey medium.
- Yeast extract Kyowa C Yeast extract Kyowa H
- Yeast extract Kyowa L Yeast extract Either Kyowa W
- Yeast Extract Takeda HR Yeast Extract Takeda SL-W or Yeast Extract JT-21A was added to prepare a medium.
- a medium supplemented with Yeast Extract for medium was prepared. The composition of the prepared medium is shown in Table 5, and the medium Nos. 1, 3, 5, 7, 9, 11 and 1-13 have MgSO 0.05%, MnSO 0.025%,
- G glucose
- C casein enzyme digest
- YEB yeast extract for medium (Batato)
- YE yeast extract for food additives
- 0 o Rain acid compound.
- (YEB) Yeast Extract (Batato) for medium
- (C) Yeast Extract Kyowa C
- (H) Yeast Extract Kyowa H
- (L) Yeast Extract Kyowa L
- (W) Yeast Extract Kyowa W
- (HR) Yeast Extract Takeda HR
- (SL-W) Yeast Extract Takeda SL-W
- (21A) Yeast Extract JT-21A
- T80 Tween80 o
- the yeast extract-added medium is a medium containing 0.3% of yeast extract Kyowa W or yeast extract Takeda HR as the yeast extract and glucose, casein enzyme-degraded product, Tween 80, and the casein enzyme-decomposed product, A medium supplemented with 0.3%, 0.6% or 0.9% of yeast extract Kyowa W or yeast extract Takeda HR was prepared.
- the composition of the prepared medium is shown in Table 6.
- G glucose
- C casein enzyme digest
- YE yeast extract for food additive
- 0 oleic acid compound.
- W yeast extract Kyowa W
- HR yeast extract Takeda HR
- T80 Tween80.
- L. acidophilus JCM11047 or L. gasseri JCM1131 was used as a test bacterium.
- the test bacterial solution that had been stored frozen was inoculated into 3 ml of 5 ml of MRS medium. Cultured in an incubator (37 ° C, 24 hr). Thereafter, the culture solution was further inoculated into 5 ml of MRS medium in an amount of 3% and cultured.
- Example 1-11 The medium described in 6 was inoculated with 3% of the bacterial solution after pre-culture of the test bacteria, stirred, and dispensed into a screw test tube. Cultivate in an incubator at 37 ° C, sample the medium at 0, 4, 8, 16, 24 hours, and measure the turbidity and pH of the medium. It was investigated.
- test examples 3% of the test bacterial solution pre-cultured in the medium was inoculated, and cultured at 37 ° C and 80 rpm using a mini jar mentor (Takasugi Corporation). In addition, the pH was maintained at 5.5 by dropping 3N_NaOH solution. The growth of the test bacteria in the medium was examined by sampling the medium over time and measuring the turbidity. [0034] Examples of tests using the lactic acid bacteria culture medium of the present invention are shown below as test examples.
- the enzyme-decomposed whey medium shown in Table 2 prepared in Example 1 was used.
- MRS medium was used.
- a 50 ml medium was inoculated with 3% of the bacterial solution of L. acidophilus JCM11047 pre-cultured by the method described in Example 7, and after stirring, 7 ml was dispensed into 7 screw-mouth test tubes. Three of them were cultured in an incubator at 37 ° C, and the turbidity and pH were measured after 0, 4, 8, 16, and 24 hours, and the viability of the test bacteria in the enzyme-degraded whey medium was examined.
- the turbidity of the three test tubes inoculated with the test bacteria was measured over time using a digital colorimeter (mini photo 10: Sangen Industrial Co., Ltd.) (measurement wavelength: 620 nm), and the three measured values Comparison was made using a value obtained by subtracting the turbidity (blank) before the start of cultivation of the average power of.
- the pH was measured over time using a pH meter (F-21 type: Horiba, Ltd.).
- the medium is a basic whey medium supplemented with glucose 1.0%, casein enzyme degradation product 0.3%, Tween 80 0.1%, and yeast extract Kyowa W 0.3% as an alternative to Yeast Extract for the medium.
- the medium supplemented with Tween 80 in Table 4 prepared in Example 4 was used.
- the MRS medium of Reference Example 1 was used.
- a 3% amount of a pre-cultured bacterial solution of L. acidophilus JCM11047 was inoculated into the MRS medium and the medium Nos. 1 and 3-5 in Table 4, and cultured in the same manner as in Test Example 1. Turbidity and pH were measured in the same manner as in Test Example 1.
- the medium supplemented with sodium oleate as an alternative to Tween 80 has high turbidity, so it was diluted 3 times with distilled water and measured, and the value obtained by doubling the turbidity was used as the result.
- the turbidity and pH were measured 16 and 24 hours after the culture using the culture media of Nos. 2, 6, and 7 in Table 4, and the growth of the test bacteria in the culture media was confirmed.
- the medium supplemented with sodium oleate as an alternative to Tween80 showed higher turbidity than the medium supplemented with Tween80.
- the turbidity was higher than that of MRS medium at 16 and 24 hours after culturing.
- the medium supplemented with sucrose oleate showed a lower turbidity than the medium supplemented with other oleate compounds.
- the medium supplemented with 0.1% or 0.2% decaglycerin monooleate showed the same degree of turbidity as the medium supplemented with Tween80. Therefore, it was shown that a medium using sodium oleate and decaglycerin monooleate as a substitute for Tween 80 has a sufficient growth promoting effect on the test bacteria.
- yeast extract-added media media Nos. 1, 3, 5, 7, 9 and 11-13 of Table 5 were used.
- 3% of the bacterial solution was inoculated after pre-culture of L. acidophilus JCM11047 or L. gasseri JCM1131, and cultured in the same manner as in Test Example 1. Turbidity was measured in the same manner as in Test Example 1. From the turbidity of these media, the viability of the test bacteria in each media was confirmed.
- Y, Kyowa C, Yeast Extract Kyowa H, Yeast Extract Kyowa L, and Y of Yeast Extract Kyowa W are added as medium Nos. 2, 4, 6, 8, and 10 in Table 5, respectively.
- the cultured medium showed higher turbidity than the medium supplemented with Yeast Extract for medium, and it was confirmed that the growth of L. acidophilus JCM11047 in each medium was promoted.
- the culture mediums Nos. 1, 3, 5, 7, 9, and 11 1-13 in Table 5 are Yeast Ex-Kyowa C, Yeast Extract Kyowa H, Yeast Extract Kyowa L, Yeast Extract Kyowa W, Yeast
- good growth of acidophilus JCM11047 was confirmed.
- yeast extract Kyowa C yeast extract Kyowa L
- yeast extract Kyowa W yeast extract Takeda HR
- yeast extract Takeda SL-W yeast extract JT-21A were added.
- the fermented medium showed turbidity equivalent to or higher than that of the medium supplemented with Yeast Extract for medium, and good growth of L. gasseri JCM1131 in the medium was confirmed.
- Yeast Extract Kyowa C Yeast Extract Kyowa H, Yeast Extract Kyowa L, Yeast Extract Kyowa W, Yeast Extract Takeda HR, Yeast Extract Takeda SL-W and Yeast Extract JT-21A are used as an alternative to Yeast Extract for medium.
- the medium was sufficient to promote growth against the test bacteria. It was shown to have fruit.
- the yeast extract-added medium for food additives shown in Table 6 prepared in Example 6 was used.
- MRS medium prepared in the same manner as in Reference Example was used.
- the yeast extract-added medium As the yeast extract-added medium, the medium No. 1-8 in Table 6 was used, and 3% of the bacterial solution after preculture of acidophilus JCM11047 was inoculated and cultured in the same manner as in Test Example 1. The turbidity was measured in the same manner as in Test Example 1. From the turbidity of these media, the viability of the test bacteria in each media was confirmed.
- the culture medium to which either Yeast Ex-Kyowa W or Yeast Extract Takeda HR is added as the medium No. 1-8 in Table 6 is either acido philus JCM11047 or L. gasseri JCM1131. Also showed high turbidity.
- the medium supplemented with 0.6% or 0.9% yeast extract showed higher turbidity than the medium supplemented with 0.3% casein enzyme degradation product and had a sufficient growth promoting effect on the test bacteria. .
- Whey medium A supplemented with glucose, casein enzyme degradation product, yeast extract Kyowa C, and Tween 80 in basic whey medium
- whey medium B supplemented with inorganic salts of MgSO and MnSO
- a bacterial solution obtained by further culturing the test bacteria pre-cultured by the method described in Reference Examples for 16 hours in MRS medium was used.
- the MRS medium and the prepared medium were inoculated with 3% of the bacterial solution of L. acidophilus JCM11047 and cultured at 37 ° C and 80 rpm using a mini jar mentor. Also,
- the 3N-NaOH solution was added dropwise to maintain the pH at 5.5.
- the medium was sampled over time, the turbidity was measured in the same manner as in Test Example 1, and the growth of the test bacteria was examined by measuring the weight of the cells after completion of the culture.
- Cultivation is completed after 24 hours, and the medium is centrifuged (3000rpm, 5 ° C, 15min) using a cooling high-speed centrifuge.
- the precipitated cells are collected, added with sterilized distilled water, and centrifuged (3000rpm). , 5 ° C, 15 min).
- the washed cell dispersion solution was dispensed into an eggplant-shaped flask, pre-frozen at -38 ° C, freeze-dried, and the cell weight was measured.
- the weight of the cells after the culture is shown in FIG. In FIG. 11, compared to the weight of the cells cultured in the MRS medium, the dry cell weight of whey medium A was 2 times or more, and that of whey medium B was 3 times or more.
- Table 8 shows the turbidity at 0, 4, 8, 16, and 24 hours after culturing and the pH at 16 and 24 hours. As shown in Fig. 12, the turbidity at 24 hours after culturing showed higher turbidity than when cultured in an incubator using a test tube. It was shown that the effect of growth of the test bacteria on the medium using whey can be further enhanced by maintaining this state.
- the medium obtained in the present invention can cultivate lactic acid bacteria in large quantities at low cost and safely by effectively using the whey that has been discarded and using an approved food additive as an additive.
- lactic acid bacteria cultured in this medium can be used for the development and production of foods, medicines and the like.
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Cited By (4)
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JP2009044983A (ja) * | 2007-08-17 | 2009-03-05 | Tohoku Univ | ガセリシンaの生産方法及び食品保存剤 |
CN103103145A (zh) * | 2011-11-11 | 2013-05-15 | 光明乳业股份有限公司 | 植物乳杆菌促生长因子及其原料组合物、制备方法和应用 |
JP2015167474A (ja) * | 2014-03-04 | 2015-09-28 | 学校法人日本大学 | ラクトバチルス属乳酸菌培養用食品グレード培地、ラクトバチルス属乳酸菌の培養方法及びラクトバチルス属乳酸菌培養物の製造方法 |
FR3108122A1 (fr) * | 2020-03-13 | 2021-09-17 | Indigo Therapeutics | Milieu de culture de qualite alimentaire |
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KR102270024B1 (ko) * | 2019-07-19 | 2021-06-30 | 유한회사 비피코리아 | 반려동물용 고체 유산균 및 이의 제조방법 |
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JP2003274851A (ja) * | 2002-03-22 | 2003-09-30 | Ajinomoto Co Inc | 液状ヨーグルト様食品の製造方法 |
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- 2005-03-17 JP JP2006511960A patent/JP4780565B2/ja not_active Expired - Fee Related
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JP2009044983A (ja) * | 2007-08-17 | 2009-03-05 | Tohoku Univ | ガセリシンaの生産方法及び食品保存剤 |
CN103103145A (zh) * | 2011-11-11 | 2013-05-15 | 光明乳业股份有限公司 | 植物乳杆菌促生长因子及其原料组合物、制备方法和应用 |
CN103103145B (zh) * | 2011-11-11 | 2015-05-27 | 光明乳业股份有限公司 | 植物乳杆菌促生长因子及其原料组合物、制备方法和应用 |
JP2015167474A (ja) * | 2014-03-04 | 2015-09-28 | 学校法人日本大学 | ラクトバチルス属乳酸菌培養用食品グレード培地、ラクトバチルス属乳酸菌の培養方法及びラクトバチルス属乳酸菌培養物の製造方法 |
FR3108122A1 (fr) * | 2020-03-13 | 2021-09-17 | Indigo Therapeutics | Milieu de culture de qualite alimentaire |
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JP4780565B2 (ja) | 2011-09-28 |
TW200533743A (en) | 2005-10-16 |
JPWO2005097972A1 (ja) | 2008-07-31 |
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