經濟部智慧財產局員工消費合作社印製 495551 A7 B7 五、發明說明（1) 【發明背景】 內包性固定化微生物經硏究証實於最小之單位體積 內可獲致最高之生物質量（Biomass)，而且其對外在環境衝 擊具有緩衝及保護的效果，故內包性微生物固定化技術應 用於廢水生物處理程序具前瞻性之開發價値。 固定化微生物系統應用於廢水處理技術之硏究，迄今 已有十餘年之歷程，目前已有許多實際應用在工業生產的 成功案例，例如高果糖糖漿、6-APA、L-氨基酸等生化產 品。一般可將固定化微生物技術區分成兩大類：第一類爲 附著性微生物之自然固定化技術；第二類爲內包性微生物 之人工固定化技術，其中藻膠酸錦（Calcium alginate)爲 使用最普遍之擔體材料之一，其他較常見的材料尙有聚乙 二醇（Ployethylene glycol，PEG)、聚乙儲醇（Polyvinyl alcohol，P VA)、鹿角藻膠（K-Carrageenan)、漬脂（&8丑1·) 、明膠（gelatin)等。 但以上材料在使用上仍存有一些缺點，包括聚合劑具 有生物毒性、機械強度不足、較難製成有利於一般連續操 作式反應器的球形顆粒、以及價格偏高等。因此，若要將 固定化技術有效應用於廢水處理與生化商品的工業製程上 ，其成功與否的關鍵，乃在於開發對微生物不具毒性、對 酵素活性不會產生干擾，而且成本低廉及具有強韌機械強 度之擔體材料。 聚乙烯亞胺（PEI)、聚乙二醇（PEG)及藻膠酸鈣 2 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） 111n.--------------^^Ti·*-------線 (請先閱讀背面之注意事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 495551 A7 ______ B7 五、發明說明（2) (Alg)等原料對生物不具毒性，在充分混合後，製形容易、 機械強度高，而且是低價量產的工業化原料。因此，非常 適合作爲固定化微生物的擔體。 【現有相關固定化微生物技術及其缺點】 近年來，歐美文獻及日本、臺灣的專利公報已公布種 種有關利用不同原料製作擔體的技術。譬如，將PVA水溶 液與微生物混合後施以冷凍真空乾燥或以冷凍回溫法進行 膠凝固定（日本專利特開昭5 7 - 1 4 1 2 9，特開昭6 1 - 1 3 9 3 8 5 ); 施以紫外線照射，形成光交聯（p h 〇 t 〇 c r 〇 s s 1 i n k i n g )結構的凝 膠方法（日本專利特開平卜4 5 43 72);另外，亦有將PVA在硼 酸一磷酸鹽中二階段成膠的方法（中華民國專利公告號 199906)。上述方法雖然可以獲得高強度的膠體以作爲固定 化微生物之擔體，但卻存有不少缺點有待改良，如在冷凍 乾燥法中，利用低溫將材料脫水至一定的含水率，步驟繁 雜、費時甚久，而且耗費能源；光交聯法則以製作薄膜爲 佳，事實上並不適合微生物固定化程序；而硼酸一磷酸鹽 二階段凝膠硬化法雖然製程簡速，而且對微生物的傷害也 較小，但其內部結構緊密，容易影響物質傳輸效果，亦是 其主要的缺失。 總而言之，上述既有技術的共同缺點有：（1 )擔體製作 費時，量產設備龐大，導致生產成本升高，生產力降低；（ 2 )低溫、真空及使用硼酸等，對微生物皆屬不利；（3 )結 構緊密，影響氣體與物質的傳輸，不利於擔體的長期使用- 3 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） II-------0^--------訂 -------線· (請先閱讀背面之注意事項再填寫本頁) A7Printed by the Employees' Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 495551 A7 B7 V. Description of the invention (1) [Background of the invention] The encapsulation of immobilized microorganisms has been researched to obtain the highest biomass in the smallest unit volume. Moreover, its external environmental impact has a buffering and protecting effect, so the immobilized microorganism immobilization technology has a prospective development price for the biological treatment of wastewater. The research on the application of immobilized microbial systems in wastewater treatment technology has more than ten years history. At present, there have been many successful cases of practical application in industrial production, such as high-fructose syrup, 6-APA, L-amino acid and other biochemical products. Generally, the technology of immobilized microorganisms can be divided into two categories: the first is the natural immobilization technology of adherent microorganisms; the second is the artificial immobilization technology of encapsulated microorganisms, of which Calcium alginate is used One of the most common carrier materials. Other common materials include Polyethylene glycol (PEG), Polyvinyl alcohol (P VA), K-Carrageenan, and grease. (&Amp; 8 ugly 1 ·), gelatin, etc. However, there are still some shortcomings in the use of the above materials, including the biological toxicity of the polymerization agent, insufficient mechanical strength, the difficulty in making spherical particles that are beneficial to the general continuous operation reactor, and the high price. Therefore, if the immobilization technology is to be effectively applied to the industrial processes of wastewater treatment and biochemical commodities, the key to its success is to develop non-toxic to microorganisms, not to interfere with enzyme activity, and low cost and strong Material for tough mechanical strength. Polyethyleneimine (PEI), Polyethylene Glycol (PEG) and Calcium Alginate 2 This paper is sized for China National Standard (CNS) A4 (210 X 297 mm) 111n .-------- ------ ^^ Ti · * ------- line (please read the precautions on the back before filling this page) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 495551 A7 ______ B7 V. Description of Invention (2) Raw materials such as (Alg) are non-toxic to living organisms. After being fully mixed, they are easy to shape, have high mechanical strength, and are industrial raw materials for mass production at low prices. Therefore, it is very suitable as a support for immobilized microorganisms. [Existing related immobilized microorganism technology and its shortcomings] In recent years, European and American literature and Japanese and Taiwanese patent gazettes have published a variety of technologies for making carriers from different raw materials. For example, the PVA aqueous solution is mixed with microorganisms and then subjected to freeze-vacuum drying or gel-fixation by freezing temperature recovery method (Japanese Patent Laid-Open Sho 5 7-1 4 1 2 9 and Japanese Patent Laid-Open Sho 6 1-1 3 9 3 8 5); a gel method of forming a photo-crosslinked (ph 〇 〇cr 〇ss 1 inking) structure by irradiating ultraviolet rays (Japanese Patent Laid-Open No. 4 5 43 72); in addition, there is also PVA in boric acid monophosphate A two-stage gelation method in salt (ROC Patent Publication No. 199906). Although the above method can obtain a high-strength colloid as a support for immobilized microorganisms, there are many shortcomings that need to be improved. For example, in the freeze-drying method, the material is dehydrated to a certain moisture content at a low temperature. It takes a long time and consumes energy. The photo-crosslinking method is better for making films, which is actually not suitable for microbial immobilization procedures. The boric acid-phosphate two-stage gel hardening method has a simple process and is less harmful to microorganisms. However, its internal structure is close and it is easy to affect the effect of material transport, which is also its main deficiency. In summary, the common disadvantages of the above-mentioned existing technologies are: (1) the time-consuming production of the load, the large-scale production equipment, leading to increased production costs and reduced productivity; (2) low temperature, vacuum, and the use of boric acid, which are not good for microorganisms; (3) The structure is tight, which affects the transmission of gases and substances, which is not conducive to the long-term use of the carrier-3 This paper size applies to China National Standard (CNS) A4 (210 X 297 mm) II ------- 0 ^ -------- Order ------- line · (Please read the notes on the back before filling this page) A7
⑽5551 五、發明說明（3) 本發明之目的即在於提出製作過程簡速，成、本低廉， 生物毒性最少，且在短時間內即可獲得耐水性、機械強度 俱局’且具優異生化活性的微生物固定化擔體，並可將其 廣泛運用在廢水處理領域和生化產業上。 [發明內容與特點】 本發明乃提供一種微生物或酵素固定化擔體之創新 製備方法，其製作過程爲：將100ml的ALG-PEG-PEI(重量 百分比分別在1〜10%之間）混合溶液加入3ml的高濃度菌液 或酵素，在充分攪拌均勻後，以滴定器滴入5〜8 % (w/v)之 C a C 1:固定液中，配合緩慢之攪拌使Alg-PEG-PEI溶液凝聚 而形成球體。其不同於其他擔體製作之處爲，擔體在固定 液中停留時間爲3〜4小時，之後以淸水淋洗2〜3分鐘，再置 入淸水中以磁石攪拌6〜9小時，以溶出擔體內的PEG，之後 即可置入4 °C冷藏箱中保存備用。在整個處理過程中，完全 未使用任何對微生物或酵素有害的物質，對微生物的活化 不會產生干擾。 本發明方法之原理與特徵在於利用高分子材料（Alg、 PEG)與帶正電的高分子電解質之交聯劑（PEI)形成網狀結 構，透過與鈣離子的交換達到強化結構的目的，使擔體在 耐水性與機械強度上都獲得相當好的成果。而微生物或酵 素則與在具兩個以上官能基的網狀·結構中以架橋結合方式 固定，在其結構與活性上幾乎不受損害。本發明之製作流 程示如第1圖。 4 $纸張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） (請先閱讀背面之注意事項再填寫本頁) 訂——‘--- 線丨拳 經濟部智慧財產局員工消費合作社印製 495551 A7 _____B7 五、發明說明（4) 本發明所採用的各原料最佳配比（以重量百分比爲計 算基礎）爲：Alg(l 〜10%)、peg(1 〜10%)、PEI(1 〜10%)。Alg (請先閱讀背面之注意事項再填寫本頁) 爲一天然多醣類，正二價之鹽類離子對其溶液具有膠化作 用，而PEI爲帶正電之局分子聚合物，其可與Aig產生電性 相吸的特性，可增強擔體的結構，至於P E G則具有良好的 親水性’在單體成形後’以水洗方式將其溶出，可以增加 擔體的通透效率。 本發明的特徵在於首先採用多種原料來製成擔體，並 以最簡單的氯化鈣溶液即可使之凝膠硬化，在接觸時間不 長、溶液毒性不高的情況下，微生物或酵素所受到的可能 傷害將減少至最低。 根據本發明所製備的擔體適合用於廢水生物處理之 微生物菌群、酵素、微生物工業及動植物細胞等之包覆固 定化技術。就微生物而言，硝化細菌、脫硝細菌、石油裂 解菌、以及活性污泥、厭氧消化污泥、甲烷化污泥等微生 物菌群；就酵素而言，例如澱粉水解酵素、纖維素酵素及 蛋白酵素等，皆是本發明擔體可行的應用對象。 經濟部智慧財產局員工消費合作社印製 【較佳實施例之說明】 實例一 準備1 4 % (重量百分比分別在1〜1 0 %之間，比例爲 4:6:4)的八4+?£0+?£1(分子量分別爲八4:198;?£0:平均 爲1900〜2000; PEI:平均爲70000)水溶液共1⑽m卜加入3ml 高濃度菌液，充分攪拌均勻，其中，高濃度菌液之濃度大 5 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） 495551 A7 B7 五、發明說明（5 ) (請先閱讀背面之注意事項再填寫本頁) 於105 cells/c.c，菌種包括「動膠菌（Zoogloea)、色桿菌 (Chromobacterium)、假單胞菌（Pseudomonas)、芽孢桿菌 (Baillus)、球衣菌（Sphaerotilus)、諾卡氏菌（Nocardia )等」，可自污水處理廠之活性污泥中取得。之後以滴定器 滴入5〜8%(w/v)CaCl2固定液中，配合緩慢的攪拌，使 Alg-PEG-PEI混合液形成直徑約3.3〜3.6mm之球形顆粒（比 重約1.0 1〜〜1.05，機械強度41.4〜48.8%)於固定液中停留 3〜4小時後，以淸水淋洗2〜3分鐘，再重新置入淸水中以磁 石攪拌6〜9小時，製成擔體135克，與生活污水5 00ml (總 氮濃度爲154mg/l，COD210mg/l)混合，置於1000ml燒杯 中，在有氧條件下進行處理。七日後單體粒徑降爲2.4〜2·8 m m，比重1 . 0 2〜1 . 0 9，機械強度3 3 . 5〜6 1 . 2，而總氮則降爲 10.2mg/l，COD 降爲 7.8mg/l。 實例二 經濟部智慧財產局員工消費合作社印製 準備14%(重量百分比分別在1〜10%之間，比例爲4·· 6·· 4) 的Alg + PEG + PEI (分子量分別爲Alg:198 ; PEG:平均爲 1900〜2000;?£1:平均爲70000)水溶液共10011111，加入41111 高濃度菌液，充分攪拌均勻，其中，高濃度菌液之濃度大 於105 cells/c.c，菌種包括「動膠菌（Zoogloea)、色桿菌 (Chromobacterium)、假單胞菌（Pseudomonas)、牙孢桿囷 (Baillus)、球衣菌（Sphaerotilus)、諾卡氏菌（Nocardia )等」，可自污水處理廠之活性污泥中取得。之後以滴定器 6 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） 經濟部智慧財產局員工消費合作社印製 495551 A7 B7 ___ 五、發明說明（6 ) 滴入5〜8%(w/v)CaCl2固定液中，配合緩慢的攪拌，使 八4-?£0-?£1混合液形成直徑約3.3〜3.6111111之球形顆粒（比 重約1.0 1〜1.05，機械強度41.4〜4 8.8%)於固定液中停留3〜4 小時後，以淸水淋洗2〜3分鐘，再重新置入淸水中以磁石 攪拌6〜9小時，製成擔體128克。於0.2 % (w/v)之人工營養液 (Nutrient broth)(成分爲：NH4NO3 ( O.lg)、Ca(N〇3) 2 ( 0.02g)、FeS04 7HO(0.01g)、（ N H 4) 2 S 0 4 ( 0 . 1 g)、⑽5551 V. Description of the invention (3) The purpose of the present invention is to propose that the production process is simple, low cost, low cost, least biological toxicity, and can obtain water resistance and mechanical strength in a short period of time, and has excellent biochemical activity. The microorganisms immobilize the carrier and can be widely used in the field of wastewater treatment and biochemical industry. [Summary and Features] The present invention provides an innovative method for preparing microorganism or enzyme immobilized support. The manufacturing process is as follows: 100ml of ALG-PEG-PEI (weight percentage is between 1 ~ 10%) mixed solution Add 3ml of high-concentration bacterial solution or enzyme, and after stirring well, drop 5 ~ 8% (w / v) C a C 1: fixed solution with a titrator, mix with slow stirring to make Alg-PEG-PEI The solution condenses to form a sphere. It is different from other carriers in that the carrier stays in the fixing solution for 3 to 4 hours, then rinses with water for 2 to 3 minutes, and then puts in water for 6 to 9 hours with magnets to stir. Dissolve the PEG in the carrier, and then put it in a 4 ° C refrigerator for storage. Throughout the treatment process, no substances harmful to microorganisms or enzymes were used, and there was no interference with the activation of microorganisms. The principle and characteristics of the method of the present invention are to use a polymer material (Alg, PEG) and a positively charged polymer electrolyte cross-linking agent (PEI) to form a network structure, and the purpose of strengthening the structure is achieved through exchange with calcium ions, so that The support has achieved quite good results in both water resistance and mechanical strength. On the other hand, microorganisms or enzymes are fixed in a bridging manner in a network structure with two or more functional groups, and their structure and activity are hardly damaged. The production process of the present invention is shown in FIG. 4 $ paper size applies to China National Standard (CNS) A4 specification (210 X 297 mm) (Please read the precautions on the back before filling this page) Order ——'--- Line 丨 Staff of the Intellectual Property Bureau of the Ministry of Economic Affairs Printed by the Consumer Cooperative 495551 A7 _____B7 V. Description of the invention (4) The optimal proportion of each raw material (based on weight percentage) used in the present invention is: Alg (l ~ 10%), peg (1 ~ 10%) , PEI (1 ~ 10%). Alg (please read the precautions on the back before filling this page) is a natural polysaccharide. Positive divalent salt ions have a gelation effect on its solution, while PEI is a positively charged local molecular polymer, which can interact with Aig has the property of attracting electricity, which can strengthen the structure of the support. As for PEG, it has good hydrophilicity. After the monomer is formed, it can be dissolved out by washing, which can increase the permeability of the support. The invention is characterized in that firstly, a variety of raw materials are used to make the support, and the simplest calcium chloride solution can be used to harden the gel. Under the condition of short contact time and low toxicity of the solution, the microorganism or enzyme The possible harm will be minimized. The carrier prepared according to the present invention is suitable for coating and fixing technology of microbial flora, enzymes, microbial industry, animal and plant cells, etc. for biological treatment of wastewater. In terms of microorganisms, nitrobacteria, denitrifying bacteria, petroleum cracking bacteria, and microbial flora such as activated sludge, anaerobic digestion sludge, methanated sludge; in terms of enzymes, such as starch hydrolysis enzymes, cellulose enzymes, and Protease and the like are all feasible application objects of the carrier of the present invention. Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs [Explanation of the preferred embodiment] Example 1 Prepare 8 4+ of 14% (weight percentages are between 1 ~ 10%, the ratio is 4: 6: 4)? £ 0 +? £ 1 (Molecular weight is 8: 198;? £ 0: average 1900 ~ 2000; PEI: average 70,000) A total of 1⑽m of aqueous solution is added to 3ml of high-concentration bacterial solution, and fully stirred, among which The concentration of the bacterial solution is large. 5 This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 495551 A7 B7 V. Description of the invention (5) (Please read the precautions on the back before filling this page) on 105 cells / cc, the strains include "Zoogloea", "Chromobacterium", "Pseudomonas", "Baillus", "Sphaerotilus", "Nocardia", etc. " It can be obtained from activated sludge of sewage treatment plant. Then drip into a 5 ~ 8% (w / v) CaCl2 fixing solution with a titrator, and mix slowly to make the Alg-PEG-PEI mixed solution form spherical particles with a diameter of about 3.3 ~ 3.6mm (specific gravity about 1.0 1 ~~ 1.05, mechanical strength 41.4 ~ 48.8%) After staying in the fixing solution for 3 ~ 4 hours, rinse with 淸 water for 2 ~ 3 minutes, then put it back into 淸 water and stir with magnet for 6 ~ 9 hours to make a carrier of 135 grams , Mixed with 500 ml of domestic sewage (total nitrogen concentration of 154 mg / l, COD 210 mg / l), placed in a 1000 ml beaker, and treated under aerobic conditions. After seven days, the particle size of the monomer decreased to 2.4 ~ 2.8mm, the specific gravity was 1.02 to 1.09, the mechanical strength was 33.5 to 61.2, and the total nitrogen was reduced to 10.2mg / l, COD Reduced to 7.8mg / l. Example 2: Employees' Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs prepared 14% of Alg + PEG + PEI (with a molecular weight of Alg: 198 for weight ratios between 1-10% and 4 ···· 4) PEG: average 1900 ~ 2000; £ 1: average 70000) A total of 10011111 aqueous solution, adding 41111 high-concentration bacterial solution, and stir well, where the concentration of high-concentration bacterial solution is greater than 105 cells / cc, and the strains include " Zoogloea, Chromobacterium, Pseudomonas, Baillus, Sphaerotilus, Nocardia, etc. can be obtained from the sewage treatment plant Obtained from activated sludge. After that, titrator 6 This paper size applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) Printed by the Consumer Cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs 495551 A7 B7 ___ 5. Description of the invention (6) Drop 5 ~ 8 % (w / v) CaCl2 fixing solution, combined with slow stirring, to make the 4-4- £ 0-? £ 1 mixture into spherical particles with a diameter of about 3.3 ~ 3.6111111 (specific gravity about 1.0 1 ~ 1.05, mechanical strength 41.4 ~ 4 8.8%) After staying in the fixing solution for 3 to 4 hours, rinse with rhenium water for 2 to 3 minutes, and then re-place into the rheological water and stir with a magnet for 6 to 9 hours to make 128 g of a support. Nutrient broth at 0.2% (w / v) (composition: NH4NO3 (O.lg), Ca (N〇3) 2 (0.02g), FeS04 7HO (0.01g), (NH 4) 2 S 0 4 (0.1 g),
MgS04 7H2O(0.1g)、K2HP04 ( 0.05g)、K2H2P04 ( 〇.〇3g) 、Natrient broth (2g)、Distilled water。）中曝氣活化五 小時，之後稱取12克擔體，置入300ml之BOD瓶內（此時瓶 內擔體塡充率爲4%)，將已曝氣達飽合之營養液塡充滿 BOD瓶。將已校正之溶氧測定儀（SVNTEX D.O meter， SD-7 0)探測棒插入BOD瓶內，以磁石攪拌器（Fargo MS-90)( 調速至4)穩定攪拌，俟溶氧値穩定時開始計錄。於第96小 時時，擔體的攝氧率仍有45.4〜49.3mg02/hr，顯示擔體的 透氧率與活性極高。 實例三 準備14%(重量百分比分別在1〜10%之間，比例爲 4:6:4)的 Alg+PEG+PEI(分子量分別爲 Alg:198; PEG:平均 爲1900〜2000; PEI:平均爲70000)水溶液共100ml，加入6ml 高濃度菌液，充分攪拌均勻，其中，高濃度菌液之濃度大 於1〇5 cells/c.c，菌種包括「動膠菌（Zoogloea)、色桿菌 7 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） 11.1-1 1---------------、訂--------* 線 (請先閱讀背面之注意事項再填寫本頁) 495551 A7 B7 五、發明說明（7 ) (Chromobacterium)、假單胞菌（Pseudomonas)、芽孢桿菌 (Baillus)、球衣菌（Sphaerotilus)、諾卡氏菌（Nocardia (請先閱讀背面之注意事項再填寫本頁) )等」，可自污水處理廠之活性污泥中取得。之後以滴定器 滴入5〜8%(w/v)CaCl2固定液中，配合緩慢的攪拌，使 八4_?£0-?£1混合液形成直徑約3.3〜3.6111111之球形顆粒（比 重約1.0 1〜1.05，機械強度41.4〜48.8%)於固定液中停留3〜4 小時後，以淸水淋洗2〜3分鐘，再重新置入淸水中以磁石 攪拌6〜9小時，製成擔體130克。取100顆擔體，置於l〇〇ml 之燒杯內，共四組，分別加入80ml生活污水（成分爲N〇3-N ：1 〜25mg/l、NH3-N : 9 〜36 mg/卜 BOD: 80 〜175 mg/1、COD :150〜3 0 0 mg/1 )，並以磁_石攪拌器穩定攪拌，每隔24小時 更新各燒杯內的污水一次，同時記錄上浮的顆粒數目，連 測1 5日後，上浮顆粒數小於1 %，顯示擔體的質傳效果極佳 〇 實例四 經濟部智慧財產局員工消費合作社印製 準備14% (重量百分比分別在1〜10%之間，比例爲4:6:4) 的Alg+PEG+PEI(分子量分別爲Alg:198; PEG:平均爲 1900〜2000; PEI:平均爲70000)水溶液共100m卜加入5ml 高濃度菌液，充分攪拌均勻，以滴定器滴入5〜8%(w/v)CaCl2 固定液中，配合緩慢的攪拌，使Alg-PEG-PEI混合液形成 直徑約3.3〜3.6mm之球形顆粒（比重約1.01〜1.05，機械強度 41.4〜48.8%)於固定液中停留3〜4小時後，以淸水淋洗2〜3 8 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） 經濟部智慧財產局員工消費合作社印製 495551 A7 B7 五、發明說明（8) 分鐘，再重新置入淸水中以磁石攪拌6〜9小時，製成擔體 1 2 5克。取4克擔體共1 2組，分別置入（〇 . 1 %、0 · 5 %、1 · 〇 ) MnS04· Η2〇、（0.1、0.5、1.〇)CuS〇4· 5Η2〇、（〇·1、Ο·5、 1.0)Al2(SO4)3· 14H20 及（0.1、〇·5、1.0)FeCl3· 6H2〇 溶液 中，靜置三日，測定各單體的機械強度變化。結果顯示’ 錳（Μη — 2)、鋁（ΑΓ3)、鐵（Fe + 3)等離子對擔體之機械強度， 隨各離子濃度之增加而增加，但銅（Cu + 2)的影響則相反， 即隨著銅離子濃度的增加單體的機械強度反而呈減弱的趨 勢。 實例五 準備14%(重量百分比分別在1〜10%之間，比例爲 4:6··4)的Alg+PEG+PEI(分子量分別爲Alg:198;PEG:平均 爲1900〜2000; PEI:平均爲70000)水溶液共100ml，加入 10 Oml高濃度菌液，充分攪拌均勻，其中，高濃度菌液之 濃度大於1〇5 cells/c.c，菌種包括「動膠菌（Zoogloea)、色 桿囷（Chromobacterium)、假單胞菌（Pseudomonas)、芽孢 桿菌（Baillus)、球衣菌（Sphaerotilus)、諾卡氏菌（Nocardia )等」，可自污水處理廠之活性污泥中取得。之後以滴定器 滴入5〜8%(w/v)CaCl2固定液中，配合緩慢的攪拌，使 Alg-PEG-PEI混合液形成直徑約3·3〜3·6mm之球形顆粒（比 重約1.01〜1.05’機械強度41.4〜48.8%)於固定液中停留3〜4 小時後，以淸水淋洗2〜3分鐘，再重新置入淸水中以磁石· 9 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公爱） -TI-----------------訂---------線 (請先閱讀背面之注意事項再填寫本頁) 495551 A7 B7 五、發明說明（9) (請先閱讀背面之注意事項再填寫本頁) 攪拌6〜9小時，製成擔體1 270克。取1 000ml燒杯7個，分別 加入擔體0、50、100、150、200、250、300克，再以新鮮 之生活污水（成分爲N〇3-N: 1〜25mg/l、NH3-N: 9〜36 mg/1 、BOD: 80 〜175 mg /卜 COD: 150 〜300 mg/1)力口入（各 1000ml) ，使各杯之擔體塡充率分別爲〇%、5%、10%、15%、20% 、25 %及3 0%。以磁石攪拌器及曝氣機對各燒杯進行等速攪 拌（lOOrprn)及等量曝氣（498cm3Air/min)，俟 3、6、9、12 小時後，停止攪拌及曝氣，測量COD、ΝΗ3_Ν、Ν03-Ν 之値，結果當水力停留時間爲9小時時，10〜30%的擔體塡 充率其C Ο D去除率均接近8 0 %，而以1 5 % ( 8 1 · 6 %去除率）與 20 % (去除率82.6)最好。而NH3_N的去除率，在塡充率25 % ，水力停留時間12小時之時，達到95.5 %爲最佳；N〇3-N的 最佳去除率爲9 7 · 8 %，條件爲塡充率1 5 %、水力停留時間1 2 小時。 實例六 經濟部智慧財產局員工消費合作社印製 準備14%(重量百分比分別在1〜10%之間，比例爲 4:6:4)的 Alg+PEG+PEI(分子量分別爲 Alg:198; PEG:平均 爲1900〜2000; PEI:平均爲70000)水溶液共100m卜加入6ml 高濃度菌液’充分攪拌均勻’其中’高濃度菌液之濃度大 於1〇5 cells/c.c，菌種包括「動膠菌（Zoogloea)、色桿菌 (Chromobacterium)、假單胞菌（Pseudomonas)、芽孢桿菌 (Baillus)、球衣菌（Sphaerotilus)、諾卡氏菌（Nocardia 10 '本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公髮) " 495551 A7 B7___ 五、發明說明（10) (請先閱讀背面之注意事項再填寫本頁) )等」，可自污水處理廠之活性污泥中取得。之後以滴定器 滴入5〜8%(w/v)CaCl2固定液中，配合緩慢的攪拌，使 Alg-PEG-PEI混合液形成直徑約3·3〜3·6mm之球形顆粒（比 重約1 .01〜1 .05，機械強度41.4〜48.8%)於固定液中停留3〜4 小時後，以淸水淋洗2〜3分鐘，再重新置入淸水中以磁石 攪拌6〜9小時，製成擔體1 2 8克。於上流式曝氣流體化床反 應槽（Upflow Aeration Fluidized Bed Reactor，UAFBR)中 塡充15%擔體，水力停留時間9小時，與國立雲林科技大學 污水處理廠（採延長式曝氣活性污泥法，水力停留時間約22 小時）進行同步比較，實驗連續進行60天，其中污水成分爲 N〇3-N: 10 〜20mg /卜 NH3-N: 15 〜30 mg/卜 BOD: 90 〜165 mg/1 、COD: 150〜300 m g/1。結果，污水廠之COD平均去除率 爲90.3%，NH3-N平均去除率爲96.1%，而N〇3-N則有累積 現象；而充塡擔體的UAFBR之COD平均去除率爲81.4% 經濟部智慧財產局員工消費合作社印製 ，NH3-N的平均去除率爲91 .4%，而N〇3-N則比污水廠減少 5 0 mg/L，顯示固定化微生物擔體在好氧條件下操作仍具有 脫氮能力，且於較短之水力停留時間內即能獲得對有機物 之良好去除效率。在連續操作60天後，擔體的外型依然完 整，富於彈性，且不易碎裂。 雖然以上已經參照實例具體說明本發明，惟該些實例 非用以限制本發明，即大凡依本發明之揭示所作的均等變 化和修飾例皆應仍屬於本發明的精神與範疇內。 11 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐） 495551 A7 _B7_ 五、發明說明（11) 【圖式之簡單說明】 第1圖係本發明之微生物或酵素固定化擔體的製造流程圖 (請先閱讀背面之注意事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 本紙張尺度適用中國國家標準（CNS)A4規格（210 X 297公釐）MgS04 7H2O (0.1g), K2HP04 (0.05g), K2H2P04 (0.03g), Natrient broth (2g), Distilled water. ) Activated for five hours under medium aeration, then weighed 12 grams of weight, put it into a 300ml BOD bottle (at this time, the weight of the weight in the bottle was 4%), and filled the aerated nutrient solution with water. BOD bottle. Insert the calibrated dissolved oxygen meter (SVNTEX DO meter, SD-7 0) probe into the BOD bottle, and stir it with a magnetic stirrer (Fargo MS-90) (adjust the speed to 4). When the dissolved oxygen is stable, Begin recording. At the 96th hour, the oxygen uptake of the carrier was still 45.4 to 49.3 mg02 / hr, indicating that the oxygen permeability and activity of the carrier were extremely high. Example 3 Prepare 14% (weight percentage between 1 ~ 10%, the ratio is 4: 6: 4) of Alg + PEG + PEI (molecular weight is Alg: 198; PEG: average 1900 ~ 2000; PEI: average A total of 100 ml of aqueous solution of 70,000) was added with 6 ml of high-concentration bacterial solution, and stirred well. The concentration of the high-concentration bacterial solution was greater than 105 cells / cc, and the strains included "Zoogloea, Chromobacterium 7 papers. Standards are applicable to China National Standard (CNS) A4 specifications (210 X 297 mm) 11.1-1 1 ---------------, order -------- * line (please (Please read the notes on the back before filling this page) 495551 A7 B7 V. Description of the Invention (7) (Chromobacterium), Pseudomonas, Baillus, Sphaerotilus, Nocardia ( Nocardia (please read the precautions on the back before filling out this page), etc. "can be obtained from the activated sludge of the sewage treatment plant. Then drip into a 5 ~ 8% (w / v) CaCl2 fixing solution with a titrator, and with slow stirring, make the 8_? £ 0-? £ 1 mixture into spherical particles with a diameter of about 3.3 ~ 3.6111111 (specific gravity about 1.0 1 ~ 1.05, mechanical strength 41.4 ~ 48.8%) After staying in the fixing solution for 3 ~ 4 hours, rinse with 淸 water for 2 ~ 3 minutes, then put it into 淸 water again and stir with magnet for 6 ~ 9 hours to make a support 130 grams. Take 100 supports and place them in 100ml beakers. There are four groups. 80ml of domestic sewage (composition: No3-N: 1 ~ 25mg / l, NH3-N: 9 ~ 36mg / bu) BOD: 80 to 175 mg / 1, COD: 150 to 300 mg / 1), and stable stirring with a magnetic stone mixer, update the sewage in each beaker every 24 hours, and record the number of particles floating up, After 15 days of continuous testing, the number of floating particles is less than 1%, which shows that the mass transfer effect of the carrier is excellent. Example 4: The consumer cooperative of the Intellectual Property Bureau of the Ministry of Economic Affairs has prepared 14% for printing (weight percentages are between 1 ~ 10%, A ratio of 4: 6: 4) of Alg + PEG + PEI (molecular weights are Alg: 198; PEG: average 1900 ~ 2000; PEI: average 70,000). A total of 100m of aqueous solution was added to 5ml of high-concentration bacterial solution, and stirred well. , Drip into a 5 ~ 8% (w / v) CaCl2 fixing solution with a titrator, and mix slowly to make the Alg-PEG-PEI mixed solution form spherical particles with a diameter of about 3.3 ~ 3.6mm (the specific gravity is about 1.01 ~ 1.05, Mechanical strength: 41.4 ~ 48.8%) After staying in the fixing solution for 3 ~ 4 hours, rinse with rinsing water for 2 ~ 3 8 This paper size is applicable to China National Standard (CNS) A4 specifications (2 10 X 297 mm) Printed by the Consumer Cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs 495551 A7 B7 V. Description of the invention (8) minutes, then put it back into the water and stir with magnets for 6 ~ 9 hours to make the carrier 1 2 5 g . A total of 12 groups of 4 grams of support were taken and placed (0.1%, 0.5%, 1.0%) MnS04 ·· 20, (0.1, 0.5, 1.0) CuS04 · 5Η2〇, ( 0.1, 0.5, 1.0) Al2 (SO4) 3. 14H20 and (0.1, 0.5, 1.0) FeCl3. 6H2O solution, left standing for three days, and measured the change in mechanical strength of each monomer. The results show that the mechanical strength of the ions such as manganese (Μη — 2), aluminum (ΑΓ3), and iron (Fe + 3) increases with the increase of the concentration of each ion, but the effect of copper (Cu + 2) is opposite. That is, as the copper ion concentration increases, the mechanical strength of the monomer tends to decrease. Example 5: Alg + PEG + PEI (molecular weight is Alg: 198; PEG: average 1900 ~ 2000; PEI: 14% (weight percentage between 1 ~ 10%, 4: 6 ·· 4)) A total of 70,000) aqueous solution was added. 100 ml of high-concentration bacterial solution was added and stirred well. The concentration of the high-concentration bacterial solution was greater than 105 cells / cc. The strains included "Zoogloea, Syringa (Chromobacterium), Pseudomonas, Baillus, Sphaerotilus, Nocardia, etc. "can be obtained from activated sludge in sewage treatment plants. Then drip into a 5 ~ 8% (w / v) CaCl2 fixing solution with a titrator, and with slow stirring, make the Alg-PEG-PEI mixed solution form spherical particles with a diameter of about 3 · 3 ~ 3 · 6mm (the specific gravity is about 1.01) ~ 1.05 'mechanical strength 41.4 ~ 48.8%) After staying in the fixing solution for 3 ~ 4 hours, rinse with 淸 water for 2 ~ 3 minutes, and then re-place it into 淸 water with magnets · 9 This paper applies the Chinese national standard (CNS ) A4 specification (210 X 297 public love) -TI ----------------- Order --------- line (Please read the precautions on the back before filling This page) 495551 A7 B7 V. Description of the invention (9) (Please read the precautions on the back before filling in this page) Stir for 6 ~ 9 hours to make 1 270 grams of support. Take 7 1 000ml beakers and add them to the carrier 0, 50, 100, 150, 200, 250, 300 grams, and then use fresh domestic sewage (composition No.3-N: 1 ~ 25mg / l, NH3-N : 9 ~ 36 mg / 1, BOD: 80 ~ 175 mg / Bu COD: 150 ~ 300 mg / 1), orally (1000ml each), so that the filling rate of each cup is 0%, 5%, 10%, 15%, 20%, 25% and 30%. Use a magnetic stirrer and aerator to perform constant-speed stirring (100rprn) and equal-aeration (498cm3Air / min) for each beaker. After 3, 6, 9, and 12 hours, stop stirring and aeration, and measure COD, ΝΗ3_Ν As a result, when the hydraulic retention time is 9 hours, the C 塡 D removal rate of the load of 10-30% is close to 80%, and the removal rate of 15% (81.6% Removal rate) and 20% (removal rate 82.6) are the best. And the removal rate of NH3_N is best when the filling rate is 25% and the hydraulic retention time is 12 hours. The best removal rate of No.3-N is 97. 8% under the condition of filling rate. 15%, hydraulic retention time 12 hours. Example 6: Employees' cooperatives of the Intellectual Property Bureau of the Ministry of Economic Affairs print 14% of Alg + PEG + PEI (with a molecular weight of Alg: 198; PEG; the ratio is 4 to 6: 4) (Average 1900 ~ 2000; PEI: average 70,000) A total of 100m aqueous solution was added to 6ml of high-concentration bacterial solution 'well stirred evenly', where the concentration of the high-concentration bacterial solution is greater than 105 cells / cc, and the bacteria species include "moving gel Zoogloea, Chromobacterium, Pseudomonas, Baillus, Sphaerotilus, Nocardia 10 'This paper is in accordance with China National Standard (CNS) A4 Specifications (210 X 297) " 495551 A7 B7___ V. Description of the invention (10) (Please read the precautions on the back before filling this page) etc. "can be obtained from the activated sludge of the sewage treatment plant. Then drip into a 5 ~ 8% (w / v) CaCl2 fixing solution with a titrator, and mix slowly with Alg-PEG-PEI mixed solution to form spherical particles with a diameter of about 3 · 3 ~ 3 · 6mm (specific gravity about 1 .01 ~ 1.05, mechanical strength 41.4 ~ 48.8%) After staying in the fixing solution for 3 ~ 4 hours, rinse with 淸 water for 2 ~ 3 minutes, and then put it into 淸 water again and stir with magnet for 6 ~ 9 hours. 1 2 8 grams. Fill 15% of the load in the Upflow Aeration Fluidized Bed Reactor (UAFBR), with a hydraulic retention time of 9 hours, and work with the National Yunlin University of Science and Technology Sewage Treatment Plant Mud method, hydraulic retention time of about 22 hours) for synchronous comparison, the experiment was carried out continuously for 60 days, in which the wastewater composition was No. 3-N: 10 ~ 20mg / Bu NH3-N: 15 ~ 30 mg / Bu BOD: 90 ~ 165 mg / 1, COD: 150 ~ 300 mg / 1. As a result, the average removal rate of COD in the sewage plant was 90.3%, the average removal rate of NH3-N was 96.1%, and the accumulation of NO3-N was cumulative. The average removal rate of COD of the UAFBR filled with the carrier was 81.4%. Economic Printed by the Consumer Cooperatives of the Ministry of Intellectual Property Bureau, the average removal rate of NH3-N is 91.4%, while No. 3-N is 50 mg / L lower than that of the sewage plant, indicating that the immobilized microbial carrier is in aerobic conditions. The lower operation still has the ability to remove nitrogen, and a good removal efficiency of organic matter can be obtained in a short hydraulic retention time. After 60 days of continuous operation, the shape of the carrier is still complete, elastic, and not easily broken. Although the present invention has been specifically described above with reference to examples, these examples are not intended to limit the present invention, that is, all equivalent changes and modifications made according to the disclosure of the present invention should still fall within the spirit and scope of the present invention. 11 This paper size applies the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 495551 A7 _B7_ V. Description of the invention (11) [Simplified description of the figure] Figure 1 is the microorganism or enzyme immobilization load of the present invention (Please read the notes on the back before filling out this page) Printed by the Intellectual Property Bureau of the Ministry of Economic Affairs, Consumer Cooperatives This paper is sized to the Chinese National Standard (CNS) A4 (210 X 297 mm)