199906 Α6 Β6 五、發明说明（ 發明背景 ί請先閱埼计面之注意事邛再填艿本頁) 活用種種天然或合成高分子材料來包復微生物完整細 胞的固定化微生物或游索技術，在i 9 8 〇年代以降，備受注 目。而且已有不少實際應用於工當生產的成功例，例如高 朵焚循衆，6 - APA ’ L -氛基狻等生化產品的生產。通常使 用於固定化擔體的代表性高分子材料爲聚丙域·醢胺 (polyacrylamide)，鹿角藤膠（K_carrageenan),裼藤膠鋼 (sodium alginate)及復脂（agar)等。聚丙烯醯胺较其他 高分子價廉’常被採用。但是，校難製成有利於一般連續 反應器的球形颗粒，而且其單體分子及聚合促進則等皆具 毒性，不利於微生物活細胞之固定化操作。鹿角藤勝雖然 成肜容路、毒性低，但是價格较高爲其主要缺點。褐藤膠 雖也1¾’格低廉’容易貌成球形，但是，在含有磷睃鼗，麵 及钟等陽離子的反應液中，膠®强度相當不捣定，甚至崩 解u另外，议;勝體的择械強度又嫌太弱，不適>合長期操 作。因此，若欲將固定化菌艘技術有效應用於生化商品的 工業化製程，汜其是最近備受注Θ的廢水處理领域，其成 功與否之闕级，乃在於對橄生物不具毒性，成本低廉及具 有强紉機械強度之拋體材料的開發。 聚 C 烯醇（Polyvinyl a]cohol·，PVA) ’ 是由醋駿 c 燦 單體經聚合，醇化而成的水溶性樹腊。P v A不具毒性甚至 到·人體亦公認無害，製形容易，板械強度高，而且是低價 童產的工常化高分子原料。因此’非常適合作爲固定化菌 體的擔體。 本紙ft尺度適用中a β家標i'MCNS) f 4規格(210X297公釐} 199906 A6 B6 Μ濟部4-央櫺f-局®:工消合作it-'p^ 五、發明說明（） 現有的PVA固定化菌體技術及缺點 近年來’歐美文廠及日本的專利公報已公佈種種有闞 利用PVA進行微生物固定化之技術。譬如將PVA水溶液與微 生物ί昆合後施以凍結真空乾燥或以冷凍回溫法進行莜勝固 定（日本專利特開昭57-14129,特開昭61-139385) 〇旄以紫 外線照射’形成光交聯（phot〇Crosslinking)結構的莜膠 方法（日本專利特開平1 -454372)。另外亦有將PVA水溶液 與微生物菌體混合液，置於狍和硼睃水溶液接觸形成架橋 結構的徒膠技術（日本專利特開昭。上述方法 雖可狸得高強度之膠體作爲固定化菌體之擔體，但卻仍有 不少缺點有待改良。在冷凍乾螈方法中，須將材料冷凍保 持在-3〇〜-8〇t：的低溫下，而後須祀燥脱水至一定的含水 率，此種冷凍-解凍-脱水的步驟，不僅费時甚久，而且手 •續繁雜，托费能源。光交聯法則大抵以製成薄膜時所利用 之方法，不利於一般的菌體固定化程序之使用。再者， PVA·硼酸法中，微生物-?“的混合物則須與硼睃接觭浸清 I2〜24小時，方能獲得相當強度的膠體，否則膠徹脆弱。 總而言之，上述既有抆術的共同缺點有，第一，固定 化程序费時長久，手績繁雜，需要大嗤的生產設備方可量 產，導致生產成本的提升，吟低生產力。第二，低溫、真 空及堋睃皆不甚適合微生物生存之環境。尤其硼睃更具毒 性，長時間的製作裎序皆可能引起微生物活性的低降。北 野清之曾提出利用PVA與微生物菌馓混合物，置於硫睃盥 溶液中接觸之方法，企圓改良上述缺點（日本專利特開昭 -4 - (諳先Μί*卄面之注意事項再琪寫本π) ·».· •打· 線· 本紙張尺度適《中ΗΒ家樣孕(CNS>T4規格(210X297公釐） Μ濟部屮央標'f局Μ工>·«*·»?合作杜卬拟 199906 五、發明说明（） 64-549〇，64乃的1)。此方法雖然有效地縮短了固定化程 序所需時間，然而所使用的凝膠溶液濃度相當高，須以 30%硫酸鈾或硫酸銨溶液進行’如果此濃度過低時顆粒 成形不易，強度亦不佳。因此，所需材料成本即校高；而 成膠過程中高i類濃度亦對橄生物生化活性甚有不良影響 〇 本發明所提出硼酸-磷皎服二陪段成膠之方法，首先 置於:3¾至狍和滚度的硼酸溶液中姐時間成形，再於3·2〇% 磷酸盟溶液中強化。成形相當容易，且磷睃盏價格低廉， 亦爲微生物能源代謝來源，不具生物毒性。此外，盟類使 用濃度低’卟低成本。而且比硫破盟法具有更桂的膠體初 性〇 本發明之目的即在於提出製作程序簡單，低成本，且 在短時間内即可獲得高耐水性、強韌機械強度，且具優異 生化活性的PVA凝膠固定化微生物或酵素，並將其廣泛應 用在廢水處理领域及生化產業上〇 發明內容與特點 本發明提供一種聚ϋ烯醇微生物成酵素固定化擔體之 改良製備方法’包含將聚G烯醉及微生物或酵素所形成之 混合物於一濃度爲3重量％至飽和的硎峻水容液中進行凝膠 化處理而產生一凝膠球體，其改久之處在於該凝膠化處理 時間爲10分鏜至2小時，該凝膠球體並接著以3 2〇重量％磷 酸或磷酸腹水溶液浸清3〇分鐘以上使其硬化而獲得聚乙烯 本纸ft尺度適用中Ββ家樣毕(CNS)甲4規格(210x297公釐） · ---- ..................................................Μ-.* .................ir...........* ...............St {請先Μ讀卄面之注意事項再填寫本頁) . 199906 Λ 6 Η 6 五、發明説明（） 醉微生物或酵素固定化擔髋。 本發明方法之原理與特微在於利用PVA分子中的氩氧 基與棚峨·分子間的離子银橋作用使之成形；而後，再將此 不甚強固的凝膠，浸清於磷酸或磷酸篮溶液中，利用PVA 與碑駿或磷睃毁間的醏化反應使膠體硬化，獲得讨水性強 ’機械強度高且幾乎不損害橄生物或酵素生化活性的固定 化逋體。本發明之製作流程可用第一囷表示。 本發明所使用的PVA，鹼化度7〇*以上，聚合度爲1〇〇〇 〜3〇00，而以鹼化度9S%以上及聚合度15〇〇〜2〇00者较爲 迤宜。聚合度遇低則膠體不稿定，過高則黏度升高不易處 理。該PVA係以一水溶液形式與該橄生物或酵素混合，合 適之PVA濃度則在10〜2〇重量％。堋酸成形時間以飽和堋酸 水溶液進行約I5〜3〇分鐘爲较佳。 磷睃或磷酸鬣水溶液之较佳濃度爲5·15重童*，浸溃 時間約需1〜2小時，即吁獲得理想成品。該磷敗盟水溶液 T使用磷酸鈉、磷酸氩餉、磷酸二氳餉、磷酸鉀、磷睃氮 鉀、麟酸二II鉀、鱗酸接、麟駿I錢或鱗駿二I铵。 本發明方法之另一較佳實施方式爲將該硼酸與磷駿或 磷酸盟水溶液混合，而同時進行該敖膠化與硬化，時間約 爲3〇分鐘至3小時，以I-2小時爲較佳0 本發明的特微在於首先在硼酸·溶液建立膠體基礎構银 ，由於接觸時間不長，使硼酸對橄生物或酵素可能的傷害 減至最低（橋本獎所提的硼酸法，須接觸15〜24小時，曰 本專利特開昭61 -100193 )。再蚩’膠體硬化過程使用具有 -6 - 本紙張尺度边用中國β家標準(CNS)規格(210x297公龙)199906 Α6 Β6 Fifth, the description of the invention (please read the precautions of the plan first and then fill in this page) use various natural or synthetic polymer materials to cover the immobilized microorganisms or micro-organism technology of complete cells of microorganisms, It has attracted attention since the fall of the 1980s. Moreover, there have been many successful examples of practical application in industrial production, such as the production of biochemical products such as Gao Duo Xun Xunzhong, 6-APA ’L-Atmosphere. The typical high-molecular materials used for immobilization of the support are polyacrylamide, polycarryenamide, K_carrageenan, sodium alginate, and agar. Polypropylene amide is often cheaper than other polymers. However, it is difficult to make spherical particles that are good for general continuous reactors, and its monomer molecules and polymerization promotion are all toxic, which is not conducive to the immobilization of living cells of microorganisms. Although the staghorn vine wins Chengrong Road and has low toxicity, its higher price is its main disadvantage. Although the rattan gum is also easy to form into a spherical shape, but in the reaction solution containing cations such as phosphoric acid, noodles and bells, the strength of the gum® is quite undisturbed, and even disintegrates. In addition, win; The body's selective strength is too weak and unsuitable for long-term operation. Therefore, if the immobilized bacteria vessel technology is to be effectively applied to the industrialization process of biochemical commodities, it is the most recently recognized Θ wastewater treatment field. The success level of its success is due to its non-toxicity to olive organisms, low cost and Development of projectile materials with strong mechanical strength. Polyvinyl a] cohol (Polyvinyl a] cohol, (PVA) ’is a water-soluble tree made from vinegar and c chan monomers by polymerization and alcoholization. P v A is not toxic or even recognized by the human body as harmless, easy to shape, high in mechanical strength, and is a low-cost industrial raw material for childbirth. Therefore, it is very suitable as a support for immobilized cells. The ft scale of this paper is suitable for a β family standard i'MCNS) f 4 specifications (210X297 mm) 199906 A6 B6 Μ Ministry of Economic Affairs 4-Central Bei f-bureau®: industrial and consumer cooperation it-'p ^ V. Description of invention () Existing PVA immobilized cell technology and shortcomings In recent years, the patent publications of Ou Meiwen Factory and Japan have published various technologies for the immobilization of microorganisms using PVA. For example, after combining PVA aqueous solution with microorganisms, freeze-drying is applied. Or freeze fixation method (Japanese Patent Laid-Open No. 57-14129, Japanese Patent Laid-Open No. 61-139385) ○ Using ultraviolet irradiation to form a photo-crosslinking (phot〇Crosslinking) structure of the lacquer method (Japanese Patent Japanese Patent Laid-Open No. 1 -454372). In addition, there is also a glue technology that puts a mixed solution of PVA aqueous solution and microbial cells in contact with roe deer and boron solution to form a bridge structure (Japanese Patent Laid-Open Patent. Although the above method can obtain high strength The colloid is used as a carrier of immobilized bacteria, but there are still many shortcomings that need to be improved. In the freeze dry salamander method, the material must be kept frozen at a low temperature of -3〇 ~ -8〇t: Dry and dehydrate to a certain water content, this kind of The steps of freezing-thawing-dehydration are not only time-consuming, but also complicated and complicated, and require energy. The photocrosslinking method is mostly based on the method used when making the film, which is not conducive to the use of general bacterial immobilization procedures . In addition, in the PVA · boric acid method, the mixture of microorganisms-? "Must be immersed in boron for I2 ~ 24 hours to obtain a very strong colloid, otherwise the glue is completely fragile. In summary, the above-mentioned existing 抆 术The common shortcomings are: first, the fixed procedure takes a long time, the work is complicated, and it requires large production equipment to mass-produce, which leads to increased production costs and low productivity. Second, low temperature, vacuum and high temperature It is not suitable for the environment where microorganisms live. Especially boron is more toxic, and the long-term production sequence may cause a decrease in the activity of microorganisms. Kitano Kiyoshi once proposed to use a mixture of PVA and microbial bacteria in a sulfur solution The method of contact to improve the above shortcomings (Japanese Patent Laid-Open Sho-4-(Precautions for the first Μί * 卄 面 ， then write a book π) Pregnancy CNS > T4 specification (210X297mm) マ テ ギ ン デ テ ム 'f Bureau M Gong > · «* ·»? Cooperation Du Xi draft 199906 V. Description of the invention () 64-549〇, 64 is 1). Although this method effectively shortens the time required for the immobilization process, the concentration of the gel solution used is quite high, and it must be performed with a 30% uranium sulfate or ammonium sulfate solution. If the concentration is too low, the particles are not easy to form and the strength is not Therefore, the required material cost is high; the high i concentration in the gel forming process also has a bad influence on the biochemical activity of olive. The method for forming the boric acid-phosphoric acid second companion segment proposed by the present invention is first set In: 3¾ to roe and roll degree of boric acid solution in the sister time to shape, and then strengthened in 3 · 20% phosphate alliance solution. It is quite easy to form, and the price of phoenix is low. It is also a source of microbial energy metabolism and is not biotoxic. In addition, the use of low concentrations of porphyrins at low cost. Moreover, it has more colloidal initial properties than the sulfur-breaking alliance method. The purpose of the present invention is to propose a simple manufacturing process, low cost, and high water resistance, strong mechanical strength, and excellent biochemical activity in a short time. PVA gel immobilized microorganisms or enzymes, and it is widely used in the field of wastewater treatment and biochemical industry. SUMMARY OF THE INVENTION AND CHARACTERISTICS The present invention provides an improved preparation method of polyenol microbial enzyme-immobilized carriers' The mixture formed by polyglycene and microorganisms or enzymes is subjected to gelation treatment in a basic aqueous solution with a concentration of 3% by weight to saturation to produce a gel sphere. The improvement is that the gelation treatment The boring time is 10 minutes to 2 hours, and the gel spheres are then immersed in 32% by weight phosphoric acid or phosphoric acid aqueous solution for more than 30 minutes to harden them to obtain polyethylene paper ft scale. CNS) A 4 specifications (210x297 mm) · ---- ......................................... ............... Μ-. * ................. ir ........... *. .............. St {Please read the attention of the noodles first Then fill the page entry). 199906 Λ 6 Η 6 V. Description of the Invention () drunk immobilized microorganism or enzyme supported hip. The principle and special feature of the method of the present invention is to use the argonoxy group in the PVA molecule and the ionic silver bridge between the molecules to shape it; then, the less strong gel is immersed in phosphoric acid or phosphoric acid In the basket solution, the colloidal hardening reaction is carried out between PVA and Beijun or phosphoric acid to harden the colloid to obtain an immobilized compound with strong mechanical strength and almost no damage to the biochemical activity of olive organisms or enzymes. The production process of the present invention can be represented by the first font. The PVA used in the present invention has a degree of alkalinity of 70 or more, a degree of polymerization of 1,000 to 3,000, and a degree of alkalinity of 9S% or more and a degree of polymerization of 1500 to 20,000 are more suitable . If the degree of polymerization is low, the colloid is not finalized, and if it is too high, the viscosity increases and it is not easy to handle. The PVA is mixed with the olive or enzyme in the form of an aqueous solution, and the appropriate PVA concentration is 10 to 20% by weight. The formation time of the bezoic acid is preferably about 15 to 30 minutes in a saturated bezoic acid aqueous solution. The preferred concentration of phosphoric acid or phosphoric acid aqueous solution is 5.15 heavy child *, and the immersion time takes about 1 to 2 hours, that is, to obtain the ideal finished product. Sodium phosphate, argonine phosphate, dihydrophosphate, potassium phosphate, potassium phosphazide, dipotassium linate, linacid acid, linjun I, or linjun di I ammonium are used as the aqueous solution T of phosphazone. Another preferred embodiment of the method of the present invention is to mix the boric acid with an aqueous solution of phosphoric acid or phosphoric acid, while simultaneously performing the gelatinization and hardening, the time is about 30 minutes to 3 hours, with I-2 hours as the comparison The best feature of the present invention is that the colloidal basic silver is first established in the boric acid solution. Since the contact time is not long, the possible damage of boric acid to olive organisms or enzymes is minimized. ~ 24 hours, Japanese Patent Laid-open Zhao 61-100193). The re-chill ’colloid hardening process uses Chinese paper standard (CNS) specifications (210x297 gong) used in this paper.
卞 JiiBian Jii
經濟部十央標準而只工消费合作杜印¾The Ministry of Economic Affairs' ten central standards and only industrial and consumer cooperation Du Yin ¾
五、發明説明（ ；i9990b 緩衝作用的鱗敗毁溶液，而且鱗元素亦爲微生物能源代謝 之必需元素’所以對微生物而言不但無害且可能具有活化 微生物之功能。 根據本發明製麟得的PVA㈣遑合使用於酵素、工 業撇生物、廢水處理微生物菌尊及動植物細胞等的包復固 定化扶術。酵素而言，譬如澱粉水解酶、纖維素酶、蛋白 酶等。橄生物而言，酒精醱酵酵母、硝化細菌、脱硝細菌 以及活性污泥、厭氣消化污泥、甲坑化污泥、脱確污泥等 黴生物菌尊。以上所述皆爲本發明PVA莜膠方法的可行應 用對象。 發明之較佳具體實施例 (請先閱讀背面之注意事項再塡寫本頁) 裝. 實施例 經濟部屮央標準局卩Η消赀合作杜印蚁 15重量％的1=以（鹼化度"％以上，聚合度2〇〇〇)水溶液 2〇g，與脱硝污泥的滾縮液（污泥滚度50g/1)20g充分搅拌 混合（脱硝污泥微生物取自實驗室生物脱氮程序中的脱硝 槽）。捋此PVA •污泥混合物滴入緩慢授：掉的飽和棚酸水溶 液中’形成直徑约的圓球膠體，並置於該溶液中2〇分 後，再取^轉移至8«㈣雜二1齡液技溃赠鐘 ，最後將㈣顆粒取出#。製成賴定⑽生物擔體 叫，與含有雜鉀及甲_主成例_人工廢水（姐 I滾度爲醇35Gppm)浪合，置於125ml的血清瓶 中，在無氧條件下進行批式脱㈤。經2小時料後人工廢 水的硝酸氮滚度降爲32pp® ° 訂· 線.V. Description of the invention (; i9990b buffering scale destruction solution, and the scale element is also an essential element for microbial energy metabolism ', so it is not only harmless to microorganisms but may also have the function of activating microorganisms. PVA ㈣ obtained by the invention It can be used in combination with enzymes, industrial skimming organisms, wastewater treatment microbes, animal and plant cells, etc. For enzymes, such as starch hydrolase, cellulase, protease, etc. For olives, alcohol Fermented yeast, nitrifying bacteria, denitrifying bacteria, activated sludge, anaerobic digestion sludge, pitted sludge, decontaminated sludge and other mildew organisms. The above are all feasible application objects of the PVA locust glue method of the present invention The preferred specific embodiment of the invention (please read the precautions on the back before writing this page) installed. Example: The Department of Economics, the Central Standards Bureau, 叩 Η 消 菀 Cooperation, Duyin Ant, 15% by weight, 1 = to (alkaline Degree "% or more, degree of polymerization 20000) 20g of aqueous solution, and 20g of denitrified sludge rolling solution (sludge rolling degree 50g / 1), fully stirred and mixed (denitrified sludge microorganisms were taken from the experiment The denitration tank in the biological denitrification program). Squeeze this PVA • Sludge mixture into the slow-slow: the saturated aqueous shed acid solution to form a spherical colloid with a diameter of about 20 minutes, and then put it in the solution for 20 minutes, then take ^ Transfer to 8 «㈣ 杂 二 1 年 時 水 技 時 紙 钟, and finally take out the ㈣ 粒 ##. Made into Lai Ding ⑽ biological support called, and containing mixed potassium and A_main example_artificial wastewater Degree is 35Gppm of alcohol) wave-coupled, placed in a 125ml serum bottle, batch desorption under anaerobic conditions. After 2 hours of feeding, the nitrate nitrogen roll of artificial wastewater is reduced to 32pp® ° Order · line.
尺度边用中國®家料(CNS)甲4W A6 B6 199906 五、發明說明（） *{請先聞讀背面之注意事項再填寫本页) 相同的固定化微生物擔體進行重復批式脱硝試驗，每 天更新相同成份的人工廢水，並測得固定化橄生物的脱硝 速率’在第七天後達0.65mg N/g gel/h，此後，一直到第 3〇天之連續操作皆維持於此一速牟，生化活性相當鴉定。 再者利用橋本獎（日本專利特開昭01-100193)所提方法製 備相同污泥含量的固定化留體，在相同條件下，進行與此 實施例同樣之實驗。批式脱硝經2小時培養後，人工廣水 的硝酸氮泼度爲82ppm，而重復批式轼驗須在天後方達 到0.55mg N/g gel/h之脱硝速率，往後持續操作亦不甚鵪 定。 實施例2 2〇重量U^JPVA(鹼化度"％以上，聚合度2〇0〇)水溶液 ，與硝化污泥的波縮液（污泥滾度3〇g/l )，以1 : 1重量比例 均勻混合（硝化污泥微生物取自實驗室生物脱氮程序中的 硝化槽〉。PVA-污泥混合物進行與實施例（1)同樣之微生物 固定化程序。 觥濟部屮央標ill-ri負工消'ff合作杜卬« 製成之橄生物固定化擔骶（粒徂爲3mm)置於操作容猜 爲10 1的生物反應器中，迷續入流含2〇0ρρω氱氮的人工麼 水（流入量3〇 Ι/day)，擔體充填率爲25%，通氣量2〇 1/min 。經1〇天連續操作後，出流水之氟*氮滚度爲9ρρω ,並有 92%的氟氮被轉換成硝酸氮。 -8 - 本紙》尺度適用中國國家標1MCNS)T4規格(210x297公釐) 199906 A6 B6 經濟部屮处標"局W工消'^合作代卬製 五、發明説明（） 實施例3 以養猪事業廢水（COD濃度15〇〇〜2〇〇〇ppm,總氮滚度 200〜3〇〇ppm)馴化達“固月的活性污泥，經離心而得滾錄 污泥溶液，以1 : 1重量比例混合]_ g重量％ p VA (驗化度9 9 %以 上，聚合度2〇〇〇)的水溶液，此均勻混合液滴入含有5重量 %硼睃與10重量％磷酸二氩鉀的混合溶液中，浸清i小時形 成3mm粒徂的圓球膠體顆粒。製成的固定化橄生物擔體， 置於與實施例（2 )同樣的生物反應器中，進行養豬事業廢 水之處理，反應器操作條件與實施例（2)相同。經2〇天連 續操作後’出流水之COD滚度降至2〇0〜3〇〇ppm，總氮濃度 亦降爲120〜l80ppm。 實紇例4 2〇重量U0VA(鹼化度"％以上，聚合度2〇〇〇)水溶液 10呂’與碑酒酵母（Sacchramyces )離心泼縮液 (細胞滾度3〇g/l)10g充分攪拌混合，PVA-酵母菌體混合物 進行與實施例（1)同樣之微生物固定化程序。毂成的固定 化細胞擔體（粒徂爲hmUSg與含有3重量％葡萄糖的培養基 isomi混合，置於三角瓶中，於3〇。〇下振燙培養，經8小時 铰’培養液之酒精產生濃度爲10.2g/1。而以相同的菌體 量，於同樣培養條件下進行游離細胞的培養，經8小時後 ’培養液中之酒精產生濃度爲10.6gA。 -9 - (請先閱讀背面之注意事項再填寫本页) *装_ •訂· .線. 本紙張尺度遴用中B國家樣孕（CNS)甲4規格(210x297公釐） i 9 9 9 0 b A6 B6 五、發明說明（） 實施例5 20重量M〇PVA(鹼化度"％以上，取合度2〇00)水溶液 10g ,與類固酵轉換細菌（Arthrobacter simplex)波縮液 (細胞滾度2〇g/i)i〇g充分攪拌混合，此混合物進行與實施 例3同樣的橄生物固定化程序。製成的固定化橄生物擔體 (粒祖約爲2mm)l5g與含有0.2g/l氩化皮質酮 (hydrocortison)的培養基l5〇ml混合，置於容積5〇Oml的 三角瓶中振盪培養，進行類固醉的△ 1 -脱氩生化反應，經 5小時培養後，有90*的基質被轉生成昝體松 (prednisolone) 〇 實紇例6 is重量％PVA(鹼化度"％以上，聚合度2〇0〇)水溶液l〇g ，與3g的異澱粉水解酶（isoamylase)酵素液及2g的/9 -澱 粉水解蜂（卢-amy 1 as e)酵素液充分混合，並以實施例（1 ) 同樣的凝膠程序進行酵素固定化。製成的固定化酵素擔體 (粒徑約2mm)15g與含有5〇g/l液化澱粉的基質溶液l5〇ml混 合，置於容積5〇〇ml的三角瓶中振盪攪拌，進行澱粉水解 反應，於反應3小時後溶液中麥芽糖濃度可達4lg/l，基質 轉揍率约爲82*. (請先閱讀背面之注意事項再填寫本资) .¾. .訂· 經濟部t处標';1,-^約工消';'合作代卬% 圖式之簡單説明 第一圖爲本發明之微生物或酵素固定化擔體的製造流 程圖 10 本纸張尺度適用中國國家梂rMCNS)T4規格（2丨0x297公釐）Standard side use China® Home Materials (CNS) A 4W A6 B6 199906 V. Description of the invention () * (Please read the precautions on the back before filling this page) Repeat the batch denitration test with the same immobilized microbial support, The artificial wastewater with the same composition is updated every day, and the denitration rate of immobilized olive organisms is measured to reach 0.65 mg N / g gel / h after the seventh day, and the continuous operation until the 30th day is maintained at this one. Su Mou, the biochemical activity is quite stable. Furthermore, the method proposed in the Hashimoto Prize (Japanese Patent Laid-Open No. 01-100193) was used to prepare immobilized deposits with the same sludge content, and the same experiments as in this example were conducted under the same conditions. After 2 hours of batch denitrification, the nitrate splash of artificial broad water is 82ppm, and the repeated batch inspection must reach a denitration rate of 0.55mg N / g gel / h after the day, and the continuous operation is not very much in the future. Quaint. Example 2 20% by weight U ^ JPVA (alkalinity degree "% or more, polymerization degree 20000) aqueous solution, and shrinkage liquid of nitrified sludge (sludge rolling degree 3〇g / l), with 1: 1 Weight ratio is uniformly mixed (the nitrification sludge microorganisms are taken from the nitrification tank in the laboratory biological denitrification procedure.) The PVA-sludge mixture is subjected to the same microorganism immobilization procedure as in Example (1). -ri negative work elimination'ff cooperation Du 卬 «The olive bio-immobilized sacral sacrum (grain size is 3mm) is placed in a bioreactor with an operating capacity of 10 1 and the continuous flow into the nitrogen containing 200ρρωω nitrogen Artificial water (inflow 3〇Ι / day), the filling rate of the support body is 25%, and the ventilation rate is 201 / min. After 10 days of continuous operation, the fluorine * nitrogen roll of the outflow water is 9ρρω, and there is 92% of the fluorine and nitrogen are converted to nitrate nitrogen. -8-This paper "standard applies to the Chinese national standard 1MCNS) T4 specification (210x297 mm) 199906 A6 B6 Ministry of Economic Affairs Standard " Bureau W Gongxiao '^ Co-generation system V. Description of the invention () Example 3 Domestication of pig farming wastewater (COD concentration of 150 to 200 ppm, total nitrogen roll of 200 to 300 ppm) for domestic activity Sludge, centrifuged to obtain a rolling sludge solution, mixed in a weight ratio of 1: 1] _ g wt% p VA (degree of inspection 99.9% or more, degree of polymerization 20000), this homogeneous mixture It was dropped into a mixed solution containing 5 wt% boron and 10 wt% potassium dihydrogen phosphate, and immersed for 1 hour to form 3mm spherical spherical colloidal particles. The prepared immobilized olive bio-support was placed in the example (2) In the same bioreactor, the pig farming wastewater was treated, and the reactor operating conditions were the same as in Example (2). After 20 days of continuous operation, the COD roll of the outflow water dropped to 200 ~ 300ppm, the total nitrogen concentration is also reduced to 120 ~ 180ppm. Example 4 2〇 weight U0VA (alkalinity "% or more, degree of polymerization 2,000〇) aqueous solution 10 L 'and Sacchramyces (Sacchramyces) 10 g of centrifugal constriction solution (cell rolling degree 3〇g / l) was fully stirred and mixed, and the PVA-yeast cell mixture was subjected to the same microbial immobilization procedure as in Example (1). The immobilized cell support (granule) Mix hmUSg with the medium isomi containing 3% by weight of glucose and place in a triangular flask at 3.0. After 8 hours of culturing, the alcohol production concentration of the culture medium is 10.2g / 1 after 8 hours. With the same cell amount, the free cells are cultured under the same culture conditions. After 8 hours, the alcohol production in the culture medium The concentration is 10.6gA. -9-(Please read the precautions on the back before filling in this page) * Packing_ • Ordering ·. Thread. This paper size is used in the B country sample pregnancy (CNS) A 4 specifications (210x297 mm ) I 9 9 9 0 b A6 B6 5. Description of the invention () Example 5 20g M〇PVA (alkalinity degree "% or more, the combined degree of 20,000) aqueous solution 10g, with steroid-converting bacteria (Arthrobacter simplex ) Shrinkage solution (cell rolling degree 2〇g / i) i〇g was fully stirred and mixed, this mixture was subjected to the same olive bio-immobilization procedure as in Example 3. 15g of the prepared immobilized olive biosupport (about 2mm in size) was mixed with 150ml of medium containing 0.2g / l hydrocortison and placed in a triangular flask with a volume of 500ml and shaken for cultivation. A quasi-drunk △ 1 -dehydrogenation biochemical reaction was carried out. After 5 hours of incubation, 90 * of the substrate was transformed into prednisolone. Example 6 is weight% PVA (degree of alkalinity "% or more, polymerization Degree 2〇〇〇) aqueous solution 10g, with 3g isoamylase (isoamylase) enzyme solution and 2g / 9- starch hydrolysis bee (Lu-amy 1 as e) enzyme solution is thoroughly mixed, and the example ( 1) Enzyme immobilization is carried out in the same gel procedure. 15g of the prepared immobilized enzyme support (particle size is about 2mm) was mixed with 150ml of the matrix solution containing 50g / l liquefied starch, placed in a triangular flask with a volume of 500ml and shaken to carry out starch hydrolysis reaction After 3 hours of reaction, the concentration of maltose in the solution can reach 4lg / l, and the matrix beat rate is about 82 *. (Please read the precautions on the back before filling in the capital). ; 1,-^ 约 工 消 ';' Cooperative representative% Simple description of the drawings The first figure is the manufacturing flow chart of the microorganism or enzyme immobilized support of the present invention. 10 The paper size is applicable to the Chinese National Committee rMCNS) T4 Specification (2 丨 0x297mm)