201021691 九、發明說明： 【發明所屬之技術領域】 本發明係與藻類之保種作業有關，尤指一種透氣式凝 膠藻種之固定化保存方法。 【先前技術】 按，藻類原種之保存，係將無其他藻類汚染之極少細 - 胞加以長期保存之作法，而大部份重要的經濟微藻，均是 ❺ 由自然環境中採集而來的，再培養及保存於實驗室中，然 而，此作法常造成藻類因長期培養於非自然環境中，而使 知藻種的族群易產生變異，因此，發展一種可長期保存微 藻，且不受污染，又不使其喪失活性的方法，是非常重要 且必須的。 此外，由於海洋藻類並沒有像陸上植物之種子具有抗 it、耐性及可休眠等特性，且藻類之生活史除了相當複雜 蠡之外，又會隨著所在環境之影響而變換形態，因此，目前 於藻類之保種及人工養瘦上皆有相當程度之困難度以及不 便生’而極需有一套能夠用以長期保存具有優良品種之讓 苗’並進而可運用於人工大量養疫之方法。 目前，應用於藻類藻源之保存方法，係以繼代養殖法、 東《法以及冷〆東乾燥法為主，然而，繼代養殖法由於相當 耗費人力及時間，且因經常需要進行移植（移種）而會有 其他生物於移植（移種）過程中混入而導致污染，則此， 201021691 目前此一作法已經較少人使用而漸漸被淘汰； 而凌結法以及冷;東乾燥法，雖較繼代養錢簡單且不 用耗費太多人力，㈣在冷錄態之下，由於微藻細胞内 將會形成冰晶’而冰晶之形成勢必會對微藻細胞 k成知傷’況且以脆弱之微藻細胞為對象，除非細心處理， 否則大多數㈣細朗會在冷錢死亡，因此在進行藻類 冷束處理時皆會對_添加抗糊，_ 峨冰㈣。㈣，财4軸考相不3 程=毒性反應與抗束效果等因素’且同時於抗束過 义備所需之_設施才得Μ成，且微_類之不 5、冷核乾㈣速率、保存時的溫度和氣體， =響到藻類冷絲之存活率，故以冷衫式保耗類藻源 其準備手段仍是相當m複雜。 “201021691 IX. Description of the invention: [Technical field to which the invention pertains] The present invention relates to the preservation operation of algae, and more particularly to an immobilized preservation method of a breathable gelatinous algae species. [Prior Art] According to the preservation of algae stocks, there are few fine-cells that are not contaminated by other algae for long-term preservation, and most important economic microalgae are collected from the natural environment. It is cultured and stored in the laboratory. However, this practice often causes algae to be cultured in an unnatural environment for a long time, and the ethnic group of the algae species is susceptible to variation. Therefore, development of a microalgae can be preserved for a long time without being contaminated. It is very important and necessary to not lose its activity. In addition, since marine algae do not have the characteristics of resistance to it, tolerance, and dormancy as seeds of land plants, and the life history of algae is not only complicated, but also changes shape according to the environment, therefore, There is a considerable degree of difficulty in the conservation of algae and artificial nourishment, and it is extremely difficult to have a set of methods that can be used for long-term preservation of seedlings with excellent varieties and can be applied to artificial large-scale quarantine. At present, the preservation method applied to the algae algae source is mainly based on the subculture method, the east method and the cold-drying method. However, the subculture method is quite labor-intensive and time-consuming, and it is often required to be transplanted ( If the other organisms are mixed in the process of transplanting (transplanting) and cause pollution, then 201021691 is currently being used by fewer people and gradually eliminated; and the knot method and the cold method; Although it is simpler to raise money than the next generation and does not require too much manpower, (4) in the cold-recorded state, ice crystals will form in the microalgae cells, and the formation of ice crystals will inevitably affect the microalgae cells. The microalgae cells are the object. Unless careful treatment, most (4) fines will die in cold money, so when the algae cold beam treatment is carried out, the anti-paste will be added, _ 峨 ( (4). (4), the 4th axis of the financial test is not 3 factors = toxic reaction and anti-beam effect and other factors - and at the same time in the anti-beam over-prepared _ facilities have to be completed, and micro-class does not 5, cold core dry (four) The rate, the temperature and the gas during storage, = the survival rate of the algae cold wire, so the preparation method of the cold-shirt type of edible algae source is still quite complicated. "
因此，有鑑於上述各種藻類保種方式之缺失後，本 發明人乃藉由從事相關研究之多年經驗，並經過不斷之: 思及實驗後，才終於有本發明之誕生。 【發明内容】 本發明之主要目的’在於提供一種透氣式凝膠藻種之 固定化保存方法，其具有可將藻類細胞予以固定化，藉以 達成長期儲存並進而保種之功效者。 本發明之再一目的，在於提供一種透氣式凝膠蒸種之 201021691 固定化保存方法，其具有製備容易且方便使用之特點者。 為達前述之目的，本發明提供一種透氣式凝膠藻種之 固定化保存方法，其步驟包含有： a ·取一欲保種之藻類細胞，並與一藻膠溶液相互混 合； b .將混合有該藻類細胞之藻膠溶液，包覆於一固態 載體上，以使該固態載體外部得形成有一藻膠層； " c ·再將形成有該藻膠層之固態載體浸泡於一氣化I弓 ❹ 溶液中’此時該藻膠層之外表面係與該氣化鈣溶液相互接 觸，並相互反應而凝結有一具有透氣性之凝膠層； d ·再將該固態載體由該氣化鈣溶液中取出，即完成 透氣式凝膠藻種之固定化保存方法。 而本發明之上述及其他目的與優點，不難從下述所選 用實施例之詳細說明與附圖中，獲得深入了解。 【實施方式】 請配合參閱第1圖及第2圖，本發明係提供一種透氣 © 式凝膠藻種之固定化保存方法，其步驟係包含有： a取欲保種之藻類細胞11 ’並與一簾膠溶液相 互混合，而於本實施例中該藻膠溶液係為褐蒸膠溶液，其 中，該褐藻膠溶液係由約1%〜i 0%之褐藻酸納與9 〇 〇/0以上之R· 〇水（或去離子水）所構成； 、 b ·將混合有該藻類細胞1 1之褐藻膠溶液，包覆於 一固態載體2 1上（於本實施例中該固態栽體係採用直和 約為6麵之破璃珠），以使該固態載體2 1於外部形成有一 201021691 褐藻膠層3 1 ; c .再將形成有該褐藻膠層3 i之固態載體2 1浸泡 於一濃度含量約為G.1M〜〇·3Μ之氯化舞⑽h)溶液 中’此時，由於該褐藻膠層3 i之外表面係與該氣化聽 液相互接觸，因此將會相互反應，而於_轉層3工之 外表面更凝結形成有一具有透氣性之凝膠層41; d ·待該固態載體2 ;[浸泡於該氣化鈣溶液中約3〜 5分鐘後’再將該固態載體2丄由該氯蝴溶液中取出即 瘳可完成本發明之透氣式凝膠藻種之固定化保存方法。 藉此，由於本發明係於該固態載體2 1 (玻璃珠）外 圍形成有該褐藻膠層3 1，並於該褐藻膠層3工外表面又 凝結形成有-具透氣性之凝膠層4 i (如附件一所示），且 該固態載體2 1係可用以支樓該褐藻膠層3工，因此利用 本發明之固定化保存方法所製成之透氣式凝膠層其外型 可相當美觀，並具有播帶找之特點，且依據其固態載體 2 i形狀同，料製造出具衫同外狀透氣式凝膠。 ' #者’由於賴轉係具有可使其營養贼水分不 巧之特點，而可使混合於該褐藻膠層3工中之藻 11可有效地被長期儲存，並進而達到保種之目的 發明藉由該褐藻膠層3 1外表面所形成之凝膠層4 =可透氣性，因此有利褐藻膠層3 i内部之空氣交換二 内，細跑11可持績4 九口作用之進灯，而不會影響藻類細胞丄工之 子 如附件二所示^ 阑％， 201021691 況且’為使本發明之透氣式凝膠藻種之固定化保存方 法，可使所欲保種之藻類細胞1 1不被污染，因此在製備 本發明之透氣式凝膠時，係可先對該褐藻膠進行一高壓高 溫之滅菌釜處理，而使後段所製備出來之透氣式凝膠，得 嫁保其藻類細胞11之純化。 ΟTherefore, in view of the above-mentioned lack of various algae conservation methods, the present inventors have finally developed the present invention by conducting many years of experience in related research and after continuous experimentation. SUMMARY OF THE INVENTION The main object of the present invention is to provide an immobilized and preserved method for a gas-permeable gelled algae having an effect of immobilizing algae cells to achieve long-term storage and further conservation. Still another object of the present invention is to provide a method for immobilization preservation of a gas permeable gel steamed product which is characterized in that it is easy to prepare and convenient to use. For the purpose of the foregoing, the present invention provides a method for immobilizing a gas-permeable gelled algae, the steps comprising: a: taking an algae cell to be preserved and mixing with a solution of algin; b. The algin solution mixed with the algae cells is coated on a solid carrier to form an algae layer on the outside of the solid carrier; " c · immersing the solid carrier formed with the alginate layer in a gasification In the solution, the outer surface of the layer of the alginate layer is in contact with the vaporized calcium solution, and reacts with each other to coagulate a gel layer having gas permeability; d. the solid carrier is further vaporized by the gasification The calcium solution is taken out, that is, the immobilized preservation method of the gas permeable gel algae is completed. The above and other objects and advantages of the present invention will become more apparent from the detailed description of the embodiments illustrated herein. [Embodiment] Referring to Figures 1 and 2, the present invention provides a method for immobilizing a gas-permeable gelatin species, the steps of which include: a: taking algal cells 11 ' The mixture is mixed with a curtain gel solution, and in the embodiment, the alginate solution is a brown steaming solution, wherein the alginate solution is from about 1% to about 0% of sodium alginate and 9 〇〇/0. The above R·hydrophobic (or deionized water) is formed; b, the alginate solution mixed with the algae cell 1 1 is coated on a solid carrier 21 (in the present embodiment, the solid-planting system) Using a straight glass and about 6 sides of the glass beads), the solid carrier 2 1 is externally formed with a 201021691 alginate layer 3 1 ; c. The solid carrier 2 1 forming the alginate layer 3 i is then immersed in a The concentration is about G.1M~〇·3Μ in the chlorination dance (10)h) solution. At this time, since the surface layer of the alginate layer 3 i is in contact with the gasified listening liquid, it will react with each other. The surface of the layer 3 is more coagulated to form a gas permeable gel layer 41; d · the solid state carrier 2; [After immersing in the vaporized calcium solution for about 3 to 5 minutes, the solid carrier 2 is removed from the chlorine butterfly solution to complete the immobilized preservation method of the gas permeable gel algae of the present invention. . Therefore, since the present invention is formed on the periphery of the solid carrier 2 1 (glass beads), the alginate layer 3 1 is formed, and on the outer surface of the alginate layer 3, a gel layer 4 having a gas permeability is formed. i (as shown in Annex 1), and the solid carrier 21 can be used to support the alginate layer, so that the breathable gel layer produced by the immobilization preservation method of the present invention can be made to have a similar appearance. The appearance is beautiful, and has the characteristics of the belt to be found, and according to the shape of the solid carrier 2 i, the material is made into a breathable gel with the outer shape of the shirt. '#者' can be effectively stored for a long time because of its characteristic that it can make its nutrition thief water unscrupulous, so that it can be effectively stored for a long time. The gel layer 4 formed by the outer surface of the alginate layer 3 is gas permeable, so that the air exchange inside the alginate layer 3 i is favorable, and the fine running 11 can achieve a driving effect of 4 and 9 functions. The seed of the algae cell is not affected, as shown in Annex II. 阑%, 201021691 and 'In order to make the immobilized preservation method of the gas permeable gel algae of the present invention, the algae cells to be preserved can be prevented. It is contaminated. Therefore, in the preparation of the gas permeable gel of the present invention, the alginate may be subjected to a high-pressure high-temperature sterilizing treatment, and the breathable gel prepared in the latter stage may be used to marry the algae cells. Purification. Ο
然於此必須特別提出說明的是，褐藻酸納（alginate ) 乃褐藻酸（alginic acid)之鹽類化合物之統稱，為一種 陰離子的結構性多醣體，其係由褐藻之細胞壁或細胞間隙 抽取出來的，褐藻酸鈉雖不溶於水，但在鹼性水溶液中將 會自然成為黏性液體（膠狀物 工邀又澡膠浴液除了可使用褐藻膠之外，亦可改由瓊 脂、角又絲喊其他麵（如紅轉（Fureellarin)、 布糊（Funoran)及紫菜（p〇rphyra)· · •等）所抽取出 之藻膠來與水溶液混合而成。 因此，由本發明所製備之透氣式凝膠，其不僅具有製 造容易’且純於冷料存法乃更為簡單、枝使用，並 不需多餘的設狀外，本發明係可提供更 且經實作後，本發明之保種方式，能在黑暗中以存 三年以上，且在長期儲 ^保存 力，且藻體數目也沒有減少;1未使_細胞喪失生殖能 則此’由以上詳細說明後可知，本發明具有可有效將 藻類細胞長期儲存以達保種之功效，且更具有=將 之特點，因此深具進步性及實 二實 已符合專躲之規定，爰㈣發㈣ 本發月實 201021691 【圖式簡單說明】 第1圖係本發明之製造流程示意圖。 第2圖係以利用本發明所製成之透氣式凝膠藻種之組 成示意圖。 附件一係利用本發明之製法所製成之透氣式凝膠藻種 之實體照片。 附件二係青海菜原生質體包埋於褐藻膠内，培養一個 月及二個月後之顯微放大照片。 ❹ 【圖式符號說明】 藻類細胞11 固態載體21 褐藻膠層3 1 凝膠層4 1However, it must be specifically stated that alginate is a general term for a salt compound of alginic acid, which is an anionic structural polysaccharide extracted from the cell wall or cell gap of brown algae. Although sodium alginate is insoluble in water, it will naturally become a viscous liquid in an alkaline aqueous solution. (In addition to the use of alginate, it can be changed from agar to agar. Silky shouting other surfaces (such as Fureellarin, Funoran and 〇rphyra), etc.), which are extracted from the aqueous solution. Therefore, the breathable material prepared by the present invention is prepared. Gel, which not only has the advantage of being easy to manufacture, but is purer than the cold storage method, is simpler to use, does not require redundant design, and the present invention can provide more and after the implementation, the invention is guaranteed. In the way, it can be stored in the dark for more than three years, and the storage capacity is stored for a long time, and the number of algae is not reduced; 1 that the _ cells are not lost in reproductive energy, as described above, the present invention has Effectively algae Long-term storage of cells to achieve the effect of conservation, and more = will be characterized, so the depth of progress and the actual two have been in line with the provisions of special hiding, 爰 (four) hair (four) this month of the real 201021691 [simple description of the map] 1 is a schematic view showing the manufacturing process of the present invention. Fig. 2 is a schematic view showing the composition of a gas permeable gel-like algae produced by the present invention. Annex 1 is a gas permeable gel algae produced by the method of the present invention. Photograph of the entity. Annex II is a photomicrograph of the protoplasts of Qinghaicai embedded in alginate and cultured for one month and two months. ❹ [Illustration of symbols] Algae cells 11 Solid carrier 21 Alginate layer 3 1 Gel layer 4 1