1307715 九、發明說明： 、 【發明所屬之技術領域】 . 本發明係關於一種分離微生物的技術，特別係關於一種 篩選可分解聚酯之微生物的培養基組成物及其方法。 【先前技術】 每年全世界所產生的塑膠廢棄物約有14〇億公噸 (Shimao, 2001，Cwrr. 12, 242-247)，所以要 如何處理塑膠廢棄物是一件刻不容緩的問題。爲了減少塑膠 • 對環境所產生的衝擊，先進國家開始對塑膠廢棄物進行更嚴 格管制並要求進行回收，同時業界也在積極發展生物可分解 (biodegradable)塑膠作為替代方案。 關於生物可分解的研究，對可分解性高分子的相關報 導，大部分是著重於酵素（Ebeling等人，1974，五wr. «/. 47, 91_97; Fukuzaki 等人，1989,五wr.尸〇/少w. 25, 1019-1026; Williams 等人，1982, Makr.心，·. 17, 1233-1246; Williams 等人，1981，五MW. 10, 5-7)或水解(Vert 等人， ® 1981，施妙⑽CT^w.如/?;?/. 5, 30-41; Vert 等人，1994,1307715 IX. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a technique for separating microorganisms, and more particularly to a medium composition for screening microorganisms capable of decomposing polyester and a method therefor. [Prior Art] There are approximately 1.4 billion metric tons of plastic waste generated worldwide each year (Shimao, 2001, Cwrr. 12, 242-247), so how to deal with plastic waste is an urgent issue. In order to reduce the impact of plastics on the environment, advanced countries have begun to tighten control of plastic waste and require recycling, and the industry is actively developing biodegradable plastics as an alternative. Regarding biodegradable research, most of the reports on decomposable polymers focus on enzymes (Ebeling et al., 1974, V. W. «/. 47, 91_97; Fukuzaki et al., 1989, V. wr. 〇/少w. 25, 1019-1026; Williams et al., 1982, Makr. Heart, ·. 17, 1233-1246; Williams et al., 1981, MW. 10, 5-7) or hydrolysis (Vert et al. , ® 1981, Shimiao (10) CT^w. such as /?;?/. 5, 30-41; Vert et al., 1994,
Biodegradable Plastics and Polymers. Elsevier Science, Amsterdam，pi 1-22)的分解，但有關微生物的分解卻是比較 少見。 對可分解性高分子（例如聚酯）的微生物之篩選，由於無 法克服高分子通常為不溶或難溶於水之瓶頸，因此目前並無 具體可行的相關研究。 【發明内容】 本發明提供一種筛選可分解聚酯微生物之培養基組成 1307715 物，其包括添加有下列式i的介面活性劑及一聚醋材作之基 本培養基： ^ CH3(CH2)nCOONa (式 I) 其中，η為6-14間之一整數。 本發明也提供一種篩選可分解聚酯微生物的方法，包括 將待篩選之微生物樣本置於添加有下列式I的介面活 性劑及一聚醋材作之基本培養基中培養： CH3(CH2)nCOONa (式 I) 籲 其中，η為6-14間之一整數；以及 觀察於上述培養基中微生物生長過程中分解圈的形成 以判斷並分離出可分解聚酯的微生物。 【實施方式】 本發明是有關於一種篩選可分解聚酯微生物之培養基 組成物，其包括添加一種介面活性劑及一聚酯材之基本培養 基。 • 根據本發明的實施例，該培養基組合物的製備包括添加 一種介面活性劑將聚酯材與基本培養基進行乳化 (emulsify)，接著再加入瓊脂(agar)以形成瓊脂培養基。 根據本發明的實施例，該介面活性劑為具有下列式I之 介面活性劑： CH3(CH2)nCOONa (式 I) 其中，η為6-14間之一整數。 根據本發明的實施例，較佳的聚酯材係選自由聚琥珀酸 二乙酯、聚己内酯以及聚羥基丁酸酯組成之群，且該聚酯材 1307715 係呈粉末形式。然而本發明所使用的聚酯材並不限於上述的 特定形式’任何能與基本培養基乳化的聚酯材種類或形式 屬於本發明的範圍。 & 根據本發明’該基本培養基為最低營養成分之基 基。根據本發明的實施例，可為含有1%碳源之: :其中-實_中，縣本培養基包含15 = 5毫克的硫酸鐵、μ亳克的硫紙1 毫克的氯⑽(㈣2)、G.5克的酵母菌抽出物、1(；克酸鋅碳7 1.0公升的蒸館水、20克的瓊脂（pH 7 2)。克的反源、 =r放該微生物包括但二似 菌種。 刀解聚酉曰或類似聚合物能力的 可分解聚酯故射菌之培養 的介面活性劑及一聚酯材 因此本發明也提供一種筛選 基組成物，其包括添加有下列式无 之基本培養基： CH3(CH2)nCOONa (式 I) 其中，η為6-14間之一整數。 另外，本發明提供一種篩選 法’包括將待篩選之微生物樣本刀解聚酯之微生物的方 性劑及一聚酯材之基本培養 基中:養添加下列式1的介面活 CH3(CH2)nCO〇Na 1307715 其中，η為6-14間之一整數；以及 觀察於上述培養基中微生物生長過程中分解圈的形成 以判斷並分離出可分解聚酯的微生物。 根據上述篩選可分解聚酯之微生物的方法，本發明方法 係用於篩選具有分解聚酯或類似聚合物能力的菌種。根據本 發明實例，所篩選的該微生物為放射菌株，包括但不限於鏈 黴菌屬（汾genus)之放射菌與小單孢菌屬 genus)之放射菌等。 根據本發明的一較佳實施例，經過熱處理的樣本在以滅 菌水適量稀釋後’可利用將樣本劃過（streak)培養基組合物 的方式接種，也可利用在無菌狀態下塗抹樣本（spread)於培 養基組合物的方式培養，接著再從培養基組合物上觀察分解 圈的形成以判斷並分離出可分解聚醋的微生物，因此任何熟 3生物學的人士應也都能理解，本發明並不限於以特定的 方式將樣本接種在上述培養基組合物。 本fill本發明更提供一種筛選可分解聚醋之放射菌的方 之基本培養基中培養：⑷^丨面活性劑及-聚醋材 CH3(CH2)nCOONa (式 I) 其中’ η為6-14間之一整數； 觀察於上述培養基中微生物 以判斷並分離出可分解㈣的放射g株。77 ·的形成 述。本發明將參考下列特定但非限制的實例作更詳盡的描 1307715 實施例： (1) 樣本之收集 在此實驗中，所採集的樣本均收集至滅菌過的毫升 管内，並在進一步處理前保存在4。(：。 (2) 篩選放射菌的培養基配方 篩選放射菌所用的培養基是澱粉·酪蛋白壤酉t (Starch-Casein Agar)，在每1公升的培養基中包括4 〇克的 澱粉、0.12克的酪蛋白、I.6克的硝酸鉀(KN〇3)、I.6克的 氯化鈉、1.6克的磷酸氫二鉀(k：2HP〇4)、40毫克的水合硫酸 鎂(MgSCU · 7H20)、16毫克的碳酸鈣(CaC〇3)、8亳克的水 合硫酸鐵(FeS〇4 · 7H20)、18.0克的瓊酯，並在培養基中添 加50微克/¾升的亞胺環己_ ( CyCi〇heximide )以抑制真菌 的生長。該製備得之培養基經121 °C滅菌15分鐘後，冷卻 至45〜50 °C ’分裝於培養皿，每一培養皿倒入15〜2〇ml，作 成培養基平面。 (3)放射菌之分離 將1毫升的底泥樣本懸浮在9毫升的滅菌水，並以50 °C 熱處理 60 分鐘（Jensen 等人，1991，五 57，1102-1108)，接著將懸浮液連續稀釋卜10000倍後，以 0.1毫升等分（每一組劃上三道）劃在殿粉酪蛋白瓊酯並於 25-28 °C培養14天，藉由重複轉移至澱粉-酪蛋白瓊酯，分 離並純化在每一樣本觀察到菌株型態的放射菌。所有分離的 菌種也可培養在不同的瓊酯培養基中，包括酵母菌抽出物-麥芽抽出物瓊酯、燕麥瓊酯、無機鹽-澱粉瓊酯及天門冬醯 1307715 胺-甘油瓊酯。 (4)篩選分解脂肪族聚酯的放射菌 把1克的脂肪族聚酯粉末溶於50毫升的二氯曱烧 (methylene chloride)中，再配製1公升的基本培養基，基本 培養基中含有0.1克的酵母菌抽出物(yeast extract)、亳克 的水合硫酸鐵、0.2克的水合硫酸鎂、1克的硫酸銨 ((NH4)2S04)、20 毫克的水合氯化鈣(caci2 · 2H20)、〇.1 克 的氯化鈉、0.5毫克的水合鉬酸鈉(Na2Mo〇4 · 2H2〇)、0.5毫 克的水合鎢酸鈉(NaW〇4 · 2H2〇)、0.6毫克的水合硫酸鎂、 50宅克的CH3(CH2)6COONa (台灣耐斯公司），再把已溶的 聚酯與基本培養基乳化’然後放置於通風櫃中加熱攪拌使二 氣甲院揮發’將18克的瓊酯加入pH 7.2的乳化培養基中， 經121 °C滅菌15分鐘，冷卻至45〜50 °C，分裝於培養皿， 每一培養皿倒入15〜20毫升，做成平面培養基，再接種放 射菌上去，並置放於30 °C的培養箱中培養。每天觀察一次， 連續觀察七天’觀察放射函菌落是否有分解圈（clear zone) 出現，並做紀錄之。 (5)放射菌分解聚酯薄膜之測試 所使用的脂肪族聚酯樣本，包括聚琥珀酸二乙酯 (poly(ethylene succinate), PES，分子量=ι.οχίο4)、聚己内酯 (poly(s-caprolactone)，PCL，分子量=6.8xl05)以及聚羥基丁 酸酯(poly(P-hydroxybutyrate)，PHB，分子量=5·4χ104)，係 購自Aldrich化學公司。利用熱壓（Linkam Hot stage)的方式 製成薄膜，將壓好的薄膜(厚度約為180微米）分別經過75% 1307715 (重量容積比）的酒精滅菌以及紫外光照射ίο分鐘，揮發完 全後的PES、PCL或PHB薄膜並稱取各1〇〇毫克，放入100 毫升的基本培養液中，再分裝1〇〇毫升至250毫升的錐形瓶 中’並以每分鐘180轉的轉速在30 °C的轉動器上保溫，隨 後滅菌並編號之。加入篩選出的放射菌（每一組實驗進行三 次），再置放於培養箱中培養。 (6)放射菌之鑑別 對培養在不同的壤S旨培養基，包括酵母菌抽出物-麥芽 抽出物瓊酯、燕麥瓊酯、無機鹽-澱粉瓊酯及天門冬醯胺-甘 油瓊酯14天的菌種進行培養特性測試。使用ISCC-NBS色 名表(Kelly, 1964，美國國家標準局國内色彩研究學會，華 盛頓特區）判斷基質菌絲體的顏色代表，在具有長距物鏡的 光學顯微鏡上觀察型態特徵並根據Hasegawa等人在1983 年提出的方法判斷細胞璧的組成(DL-與LL-二胺基庚二酸 異構物（diaminopimelic acid isomer), A2pm)。在具有 0.1 毫 升之6N鹽酸的冷;東管（Evergreen Scientific)内放入一個或二 個菌落，冷凍管以121 °C滅菌15分鐘的方式加熱，冷卻後 將1微升的水解產物放置在纖維素薄盤（microcrystalline cellulose f，東京化成株式會社），並在同一薄盤上另點上1 微升的0.01摩爾的DL-A2pm作為標準。在甲醇-蒸顧水-6N 鹽酸-吼咬(pyridine)(80:26:4:10容積/容積比）的溶劑系統上 對薄盤進行3至4小時的顯影，並待薄盤乾後以二氫三酮喷 灑劑(Ninhydrin Spray Reagent)喷灑並以 100 °C 加熱 5 分 鐘，而A2pm的點會呈現黃綠色。全細胞糖的分析步驟大致 11 1307715 與A2pm的分析步驟相同，只是水解與顯影液分別為Decomposition of Biodegradable Plastics and Polymers. Elsevier Science, Amsterdam, pi 1-22), but the decomposition of microorganisms is relatively rare. Screening of microorganisms for decomposable polymers (e.g., polyester) has not been possible to overcome the bottleneck in which the polymer is generally insoluble or poorly soluble in water. SUMMARY OF THE INVENTION The present invention provides a medium composition 1307715 for screening a decomposable polyester microorganism, which comprises a surfactant added with a surfactant of the following formula i and a polyacetate as a basic medium: ^ CH3(CH2)nCOONa (formula I) wherein η is an integer between 6 and 14. The invention also provides a method for screening a decomposable polyester microorganism, comprising culturing a sample of the microorganism to be screened in a basic medium supplemented with the following formula I and a polyacetate: CH3(CH2)nCOONa ( Formula I) wherein η is an integer of from 6 to 14; and that the formation of the decomposition ring during the growth of the microorganism in the above medium is observed to judge and separate the microorganism capable of decomposing the polyester. [Embodiment] The present invention relates to a medium composition for screening a decomposable polyester microorganism, which comprises adding a surfactant and a basic medium of a polyester material. • According to an embodiment of the invention, the preparation of the medium composition comprises the addition of an interfacial agent to emulsify the polyester material with the minimal medium, followed by the addition of agar to form an agar medium. According to an embodiment of the invention, the interfacial surfactant is a surfactant having the following formula I: CH3(CH2)nCOONa (Formula I) wherein η is an integer between 6 and 14. According to an embodiment of the present invention, a preferred polyester material is selected from the group consisting of polyethylene polysuccinate, polycaprolactone, and polyhydroxybutyrate, and the polyester material 1307715 is in powder form. However, the polyester material used in the present invention is not limited to the specific form described above. Any kind or form of the polyester material which can be emulsified with the basic medium is within the scope of the present invention. & According to the present invention, the minimal medium is the base of the lowest nutrient component. According to an embodiment of the present invention, it may be a 1% carbon source: wherein, the medium of the county contains 15 = 5 mg of iron sulfate, 1 μg of sulfur paper, 1 mg of chlorine (10) ((4) 2), G. 5 grams of yeast extract, 1 (; zinc citrate 7 1.0 liter of steamed water, 20 grams of agar (pH 7 2). The source of grams, =r put the microorganisms including but the same bacteria An intercalating agent for culturing a polycondensation or polymer-like decomposable polyester bacterium and a polyester material. The present invention therefore also provides a screening composition comprising the following formula Medium: CH3(CH2)nCOONa (Formula I) wherein η is an integer between 6 and 14. In addition, the present invention provides a screening method comprising a microbial agent for sterilizing a microorganism of a microorganism sample to be screened and In a basic medium of a polyester material: the interface active CH3(CH2)nCO〇Na 1307715 of the following formula 1 is added, wherein η is an integer of 6-14; and the decomposition circle is observed during the growth of the microorganism in the above medium. Microorganisms formed to judge and separate the decomposable polyester. A method of microorganisms of polyester, the method of the invention is for screening strains having the ability to decompose polyester or similar polymers. According to an embodiment of the invention, the microorganisms screened are radioactive strains, including but not limited to Streptomyces (汾Radiation bacteria of genus) and radiobacteria of Micromonospora genus. According to a preferred embodiment of the present invention, the heat-treated sample may be inoculated by stabilizing the medium composition after being diluted with sterilized water, or may be spread by sterilizing the sample. The medium is cultured in the form of a medium composition, and then the formation of the decomposition ring is observed from the medium composition to judge and separate the microorganism capable of decomposing the vinegar, so that any person skilled in the biology should understand that the present invention does not It is limited to seeding the sample in the above medium composition in a specific manner. The present invention further provides a method for screening a basic medium for decomposing the radiobacteria of the vinegar: (4) a surfactant and a polyacetate CH3(CH2)nCOONa (Formula I) wherein 'η is 6- One of 14 integers; the microorganisms in the above medium were observed to determine and isolate the radioactive g strain which can be decomposed (4). The formation of 77 ·. The invention will be described in more detail with reference to the following specific but non-limiting examples. 1307715 Examples: (1) Collection of samples In this experiment, the collected samples were collected into sterilized milliliter tubes and stored before further processing. In; 4. (:) (2) Screening the medium of the radiobacteria The medium used for screening the radiobacteria is Starch-Casein Agar, which includes 4 g of starch and 0.12 g per 1 liter of medium. Casein, I.6 g of potassium nitrate (KN〇3), 1.6 g of sodium chloride, 1.6 g of dipotassium hydrogen phosphate (k: 2HP〇4), 40 mg of magnesium sulfate hydrate (MgSCU · 7H20 ), 16 mg of calcium carbonate (CaC〇3), 8 g of hydrated iron sulphate (FeS〇4 · 7H20), 18.0 g of agar, and 50 μg/3⁄4 liter of imine cyclohexane in the medium. (CyCi〇heximide) to inhibit the growth of fungi. The prepared medium was sterilized at 121 °C for 15 minutes, then cooled to 45~50 °C 'packed in culture dishes, and each culture dish was poured into 15~2〇ml. Prepare the medium plane. (3) Separation of radiobacteria 1 ml of the sediment sample was suspended in 9 ml of sterilized water and heat-treated at 50 °C for 60 minutes (Jensen et al., 1991, 5, 57, 1102-1108). Then, the suspension was continuously diluted 10,000 times, and then divided into 0.1 ml aliquots (three in each group) in the case powder casein agar and at 25-28 ° C. After 14 days, by repeating the transfer to starch-casein agar, the strains of the strains were observed and purified in each sample. All the isolated strains could also be cultured in different agar medium, including yeast. Bacterial extract - malt extract agar, oat agar, inorganic salt - starch agar and aspartame 1307715 amine - glycerol agar. (4) Screening of radioactive bacteria decomposing aliphatic polyester 1 gram of aliphatic The polyester powder is dissolved in 50 ml of methylene chloride, and 1 liter of the basic medium is further prepared. The basic medium contains 0.1 g of yeast extract, gram of hydrated iron sulfate, 0.2 Grams of hydrated magnesium sulfate, 1 gram of ammonium sulphate ((NH4)2S04), 20 mg of hydrated calcium chloride (caci2 · 2H20), 1.1 gram of sodium chloride, 0.5 mg of sodium hydrated molybdate (Na2Mo〇) 4 · 2H2 〇), 0.5 mg of sodium hydrated sodium sulphate (NaW 〇 4 · 2H2 〇), 0.6 mg of magnesium sulfate hydrate, 50 oz of CH3 (CH2) 6 COONa (Taiwan Nes), and then dissolved The polyester is emulsified with the basic medium' and then placed in a fume hood to heat and stir to make the second gas Volatilization' Add 18g of agar to the emulsified medium of pH 7.2, sterilize at 121 °C for 15 minutes, cool to 45~50 °C, dispense into the culture dish, pour 15~20 ml into each dish, do The medium was plated, and then the radiobacteria were inoculated and placed in an incubator at 30 °C. Observe once a day and observe for seven consecutive days. Observe whether there is a clear zone in the radioactive colonies and make a record. (5) Samples of aliphatic polyester used in the test of radiobacteria decomposition polyester film, including poly(ethylene succinate, PES, molecular weight = ι.οχίο4), polycaprolactone (poly ( S-caprolactone), PCL, molecular weight = 6.8 x 105) and poly(P-hydroxybutyrate, PHB, molecular weight = 5.4 χ 104) were purchased from Aldrich Chemical Company. The film was formed by a hot press (Linkam Hot stage), and the pressed film (thickness of about 180 μm) was subjected to alcohol sterilization of 75% 1307715 (weight-to-volume ratio) and ultraviolet light for ίο minutes, and the volatilization was completed. PES, PCL or PHB film and weigh 1 mg each, put it into 100 ml of basic medium, and then dispense 1 ml to 250 ml Erlenmeyer flasks and rotate at 180 rpm. Incubate on a 30 °C rotator, then sterilize and number. The selected radiobacteria (three times in each set of experiments) were added and placed in an incubator for cultivation. (6) Identification of radiobacteria for culture in different soils, including yeast extracts - malt extract agar, oat agar, inorganic salt - starch agar and aspartame - glycerol agar 14 The strains of the day were tested for culture characteristics. The ISCC-NBS color name table (Kelly, 1964, National Color Institute of China National Color Research Institute, Washington, DC) was used to judge the color representative of the matrix mycelium, and the shape characteristics were observed on an optical microscope with a long-distance objective lens according to Hasegawa et al. The method proposed by the human in 1983 determines the composition of the cell ( (DL- and LL-diaminopimelic acid isomer, A2pm). One or two colonies were placed in a cold tube with 0.1 mL of 6N hydrochloric acid; the tube was sterilized at 121 °C for 15 minutes, and 1 μl of the hydrolyzate was placed in the fiber after cooling. A microcrystalline cellulose f (Tokyo Chemical Co., Ltd.) was placed on the same thin plate with 1 μL of 0.01 mol DL-A2pm as a standard. The thin disk was developed on a solvent system of methanol-hydrogen-6N hydrochloric acid-pyridine (80:26:4:10 volume/volume ratio) for 3 to 4 hours, and after the thin plate was dried, Spray with Ninhydrin Spray Reagent and heat at 100 °C for 5 minutes, while the A2pm dots will appear yellow-green. The whole cell sugar analysis step is roughly 11 1307715 and the A2pm analysis step is the same, except that the hydrolysis and developer are respectively
0.25N 的鹽酸與n_丁醇'蒸餾水-吡啶曱苯(toluene)(10:6:6:l容積/ 谷積比）而喷灑劑為酸笨胺醋（aeid⑽⑴狀phthalate)。標準糖 溶液包含了各為1%濃度的半乳糖(galact〇se)、葡萄糖、甘露 糖（mannose)、樹膠駿糖（arabin〇se)、木糖（xyi〇se)及核糖 (ribose) ° (7) 基質之利用試驗 =使用最低培養基製備含有 1%碳源之基本培養基，該 最低培養基包含丨.5克的磷酸氫二鉀、1.0克的硫酸銨、5.0 克的鱗酸二氫鉀、〇.5克的氯化鉀、5毫克的硫酸鐵、1.6毫 克的硫酸猛、M毫克的硫酸鋅、2.0毫克的氯化鈷(CoCl2)、 0.5克的酵母菌抽出物、1〇克的碳源、1〇公升的蒸餾水、2〇 克的瓊脂（pH 7.2)。其中使用之碳源包括吐溫20 (Tween 20)、甲叛基纖維素(carb〇Xy methyl cellulose (CMC))、樹膠 質（xylan)、殿粉及路蛋白。 (8) 水中細菌計數—抹盤方式（總活菌計數） 以1克的底泥樣本與9毫升的滅菌水完全混合並放置 30分鐘’接著再將1毫升的底泥樣本加入9毫升的滅菌水 以產生連續稀釋’把〇.2毫升等分的稀釋液以無菌操作方式 塗抹在瓊脂盤，並將瓊脂盤放置在30 °C培養48小時。 籁選試驗： 收集河床底泥或底泥懸浮液樣本，將該樣本以5〇 熱 處理60分鐘後’以上述培養方式培養在瓊酯培養基。 由所收集的樣本共篩選出305個放射菌株，所觀察到的 120.25 N hydrochloric acid and n-butanol 'distilled water-toluene (10:6:6:1 volume / grain ratio) and the spray was acid albino vinegar (aeid (10) (1) phthalate). The standard sugar solution contains 1% concentration of galactose, glucose, mannose, arabin〇se, xylose and ribose ° ( 7) Substrate utilization test = Preparation of a minimal medium containing 1% carbon source using a minimum medium containing 0.5 g of dipotassium hydrogen phosphate, 1.0 g of ammonium sulfate, 5.0 g of potassium dihydrogen sulfate, cesium .5 g of potassium chloride, 5 mg of ferric sulfate, 1.6 mg of sulfuric acid, M mg of zinc sulfate, 2.0 mg of cobalt chloride (CoCl 2 ), 0.5 g of yeast extract, 1 g of carbon source 1 liter of distilled water, 2 grams of agar (pH 7.2). Among the carbon sources used are Tween 20, carb Xy methyl cellulose (CMC), xylan, temple powder and road protein. (8) Bacterial count in water - wipe method (total live count) Completely mix 1 gram of sediment sample with 9 ml of sterilized water for 30 minutes' then add 1 ml of sediment sample to 9 ml of sterilized The water was applied as a serial dilution of '2 ml aliquots of the dilution to an agar plate aseptically, and the agar plate was placed at 30 ° C for 48 hours. Selection test: A sample of river bed sediment or sediment suspension was collected, and the sample was heat treated with 5 Torr for 60 minutes, and then cultured in agar medium in the above culture manner. A total of 305 radioactive strains were screened from the collected samples, and the observed 12
1307715 放射菌菌落呈現出特定似皮革、粉末或棉的乾燥表面具有粗 糙邊緣或不規則型。在底泥懸浮樣本中所發現的總微生物族 群為0.71至8.71xl04/亳升，而在底泥樣本中則為113至 56.1xl〇4/克。 接著’利用分解圈的方法從所分離出放射菌株中篩選具 分解聚合物能力的放射菌株。在瓊脂盤上形成菌落7天後， 在包含可分解聚合物之微生物的菌落周圍形成圓形分解 圈。結果顯示135個分離菌株為ρηβ-分解者（佔總數的 44.2%) ’ 83個分離菌株為PCL_分解者(佔總數的44 2%)， 而64個分離菌株能分離pES。因此，能分解pHB的分離菌 株明顯比能分解PCL或PES的分離菌株更多。在135個分 離菌株中，有46株也可以分解PCL，39株可分解PES，而 有12株是可分解PHB、PCL及PES。 在基質利用試驗中，上述12株放射菌均可以使用酪蛋 白、曱羧基纖維素及澱粉作為其碳源，而除了分離菌株 TH-11外’其餘11種也可使用吐溫20。分析上述篩選分離 出菌株的型態與生理特性，如以下表1所示，多數的分離菌 株屬於鏈黴菌屬（你^/^0所；^1^611113)(佔總數的91.9%)與小 单抱菌屬genus)(佔總數的 8.1%)。 表1 菌種 百分比（％) 鏈黴菌屬 91.9 小單抱菌屬 8.1 13 1307715 熟諳本項技藝之人士應即暸解，可對前述各具體實施例 進行修改而不致悖離其廣義發明概念。從而應暸解本發明並 不限於本揭之各項特定具體實施例，而係為涵蓋歸屬如後載 申請專利範圍中所定義之本發明精神及範_的修改項目。1307715 Radiation colonies exhibit a rough surface or irregular shape on a dry surface that is specific to leather, powder or cotton. The total microbial population found in the sediment sample was 0.71 to 8.71 x 104 / liter, while in the sediment sample it was 113 to 56.1 x 〇 4 / gram. Next, the radioactive strain capable of decomposing the polymer was screened from the isolated radioactive strain by a method of decomposing the circle. After colonies were formed on the agar plate for 7 days, a circular decomposition ring was formed around the colony containing the microorganism capable of decomposing the polymer. The results showed that 135 isolates were ρηβ-decomposers (44.2% of the total) ‘ 83 isolates were PCL_decomposers (44 2% of the total), while 64 isolates were able to isolate pES. Therefore, the isolated strain capable of decomposing pHB is significantly more than the isolated strain capable of decomposing PCL or PES. Among the 135 isolated strains, 46 can also decompose PCL, 39 can decompose PES, and 12 can decompose PHB, PCL and PES. In the substrate utilization test, the above 12 strains of the above-mentioned radiobacteria can use casein, stearyl cellulose and starch as their carbon sources, and Tween 20 can be used in addition to the isolate strain TH-11. The type and physiological characteristics of the above strains were analyzed and analyzed, as shown in Table 1 below. Most of the isolated strains belonged to the genus Streptomyces (you ^/^0; ^1^611113) (91.9% of the total) and small Genus genus) (8.1% of the total). Table 1 Strain percentage (%) Streptomyces 91.9 Pseudomonas 8.1 13 1307715 Those skilled in the art should understand that the foregoing specific embodiments may be modified without departing from the broad inventive concept. It is understood that the invention is not limited to the specific embodiments disclosed herein, but is intended to cover the modifications of the invention and the scope of the invention as defined in the appended claims.
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