201213546 六、發明說明： 【發明所屬之技術領域】 [0001] 本發明係有關一種以微生物轉化藻類醣質取得生質 原油之方法，該方法利用預定菌體的生物特性，來對藻 類體内的醣質進行生物轉換作用生產出生質原油，以取 得生質原油做為能源用油的製造來源。 [0002] 【先前技術·】 隨著全球經濟發展和人口快速增加，人類對於食用 〇 〇 油、能源石油或柴油等油脂的需求亦隨之增加，而食用 油的提煉來源主要以植物、動物脂肪為主，而能源用石 油或柴油則必須透過開採來取得，但由於需求甚大，目 前依靠植物、動物脂肪已無法滿足人類的需求，此外， 由於全球各生態物種均飽受威脅，不論是動物的養殖所 帶來環境的更迅速惡化或者是獵捕野生動物而威脅野生 動物的生存皆不是目前可行的解決方法，亦即，全球各 地的能源油礦的存量也--傳出即將用盡的警訊，故如 何節省能源以及如何尋求新的管道來取得能源已成為人 類當前最需探討研究的領域。 而目前所知，已有相當多的技術係針對於藻類中取 得油脂，而由於藻類富含有蛋白質、脂肪、藻膠以及纖 維素、多醣體甚至有微量元素和礦物質，亦即，微藻具 有葉綠素等光合器官，可有效地利用太陽能通過光合作 用將Η 2〇、C 0 2和無機鹽轉化為有機化合物，因此 可藉由微藻吸收C◦2來趨緩溫室效應，而微藻的繁殖 099132365 方式係以二分裂式為主，細胞週期較短，也易於進行大 規模培養，故在藻類g中獲得所需的物質與能源，已逐 表單編號A0101 第3頁/共13頁 0992056705-0 201213546 漸發展成為一項生物科技重要的研究領域。 而一般藻類體内含油量極低，其略僅有1 0 %以下 ，甚者僅有2%— 3%，只是在藻類培養的過程中，目 前會藉由減氮的方式或亦透過培養基來讓藻類體内所含 的油脂增加，俾使培養後的藻類可提供更多的油脂，但 也因此，延緩了在整體的培養以及後續的作用、萃取流 程，反而使其效率降低，且該增加了一道手續，成本也 因而增加。 有鑑於上述缺失弊端，本發明人認為其有待改正之 必要，遂以其從事相關產品設計製造之多年經驗，及其 一貫秉持具有之優良設計理念，針對以上不良處加以研 究創作，在經過不斷的努力後，終乃推出本發明以微生 物轉化藻類醣質取得生質原油之方法，其以更正優良之 技術以提升產業科技之功效。 【發明内容】 [0003] 099132365 本發明以微生物轉化藻類醣質取得生質原油之方法 ，主要係利用預定菌體的生物特性來有效將藻類體内的 物質進行轉換成油脂，如將纖維素分解並生成短鏈脂肪 酸，藉以提升產油流程的整體效率，來做為解決目前地 球能源快速耗盡的危機的創作理念者。 爲符合前揭之創作目的，本發明以微生物轉化藻類 醣質取得生質原油之方法，主要包含：藻類製備、添加 作用菌體、生物轉換作用、取得產物等步驟，該係在藻 類製備的過程中，省去一般會藉由減氮或亦透過培養基 的方式來讓藻類體内所含的油脂增加的培養流程，其係 因為該方式雖可在藻類中獲得較多量的油脂，但作業時 表單編號A0101 第4頁/共13頁 0992056705-0 201213546201213546 VI. Description of the Invention: [Technical Field of the Invention] [0001] The present invention relates to a method for obtaining biomass crude oil by microbial conversion of algal saccharides, which utilizes the biological characteristics of predetermined cells to treat algae Glucose is biotransformed to produce raw crude oil to obtain raw crude oil as a source of energy for oil production. [0002] [Previous Technology·] With the development of the global economy and the rapid increase of population, the demand for oils such as oyster sauce, energy oil or diesel oil has increased, and the source of edible oil is mainly plant and animal fat. Mainly, energy or oil or diesel must be obtained through mining. However, due to the high demand, the current dependence on plants and animal fats can no longer meet the needs of human beings. In addition, because all the ecological species in the world are threatened, whether it is animals or not. The more rapid deterioration of the environment brought about by breeding or the hunting of wild animals and the threat to the survival of wild animals are not currently feasible solutions, that is, the stock of energy oil mines around the world - the warning that will be exhausted Therefore, how to save energy and how to find new pipelines to obtain energy has become the most important area for human research. As far as we know, there are quite a few technical departments that aim to obtain oils and fats in algae, and because algae are rich in protein, fat, algin and cellulose, polysaccharides and even trace elements and minerals, ie, microalgae It has photosynthetic organs such as chlorophyll, which can effectively convert Η 2〇, C 0 2 and inorganic salts into organic compounds through photosynthesis by solar energy. Therefore, microalgae can absorb C◦2 to slow down the greenhouse effect, while microalgae Breeding 099132365 is mainly composed of two divisions, with a short cell cycle and easy to carry out large-scale cultivation. Therefore, the required substances and energy are obtained in algae g, which has been numbered A0101, page 3/13, 0992056705- 0 201213546 Developed into an important research area of biotechnology. In general, algae have very low oil content, which is only slightly less than 10%, and even only 2% to 3%. However, in the process of algae cultivation, it is currently reduced by nitrogen or medium. In order to increase the amount of oil contained in the algae, so that the cultured algae can provide more oil, but also delay the overall cultivation and subsequent action, extraction process, but reduce its efficiency, and A formality has been added and costs have increased. In view of the above-mentioned shortcomings, the inventor believes that it is necessary to correct it, and that it has been engaged in the design and manufacture of related products for many years, and has always adhered to the excellent design concept, and researched and created the above disadvantages. After the effort, the present invention finally introduced a method for obtaining biomass crude oil by microbial conversion of algal saccharides, which is to improve the technology of the industry by correcting the excellent technology. SUMMARY OF THE INVENTION [0003] 099132365 The present invention is a method for microbial conversion of algal saccharide to obtain raw crude oil, mainly by utilizing the biological characteristics of the predetermined bacteria to effectively convert the substances in the algae into oil, such as decomposing cellulose. And the production of short-chain fatty acids, in order to improve the overall efficiency of the oil production process, as a creative concept to solve the current global energy shortage crisis. In order to meet the purpose of the prior art, the method for obtaining biomass crude oil by microbial transformation of algae saccharide comprises the steps of: preparation of algae, addition of bacteria, biotransformation, product acquisition, etc., in the process of algae preparation In this way, the cultivation process that generally increases the oil contained in the algae by reducing nitrogen or also passing through the medium is omitted, because this method can obtain a larger amount of oil in the algae, but the operation form No. A0101 Page 4 of 13 Page 0992056705-0 201213546
[0004] Ο 效相對較長，而唯有在實施流程中降低變數的發生才能 達到最高的實用功效，更詳盡的說明就是手續流程越多 ，則增加了更多的失敗機會（即變數），而本發明在藻 類製備後直接透過藻類與菌體的生物體之間的自然轉換 作用來生產生質原油，而藻類體内約有2 0〜5 0 %以 上係為纖維素、藻膠以及多醣體等物質，故在其物質經 由轉換後的生質原油量係可有效達到較高於以往技術來 提高油脂量，此提升其整體流程的效率，且該生質原油 同時也提供了人類所需求的能源用油，藉以達到環保、 節能等良益功效並符合產業利用性者。 【實施方式】 為能更清楚表達本發明之技術重點以及為能達成所 運用的技術手段，於後述配合圖示做更詳細的說明： 本發明係有關一種以微生物轉化藻類醣質取得生質 原油之方法，〔請參閱第一圖〕主要包含： 藻類製備（1)，係將預定藻類進行規模性的培養 ，並於實施時適量地置入在反應器中，在本發明中所使 用的藻類係以非油性藻為主，而藻類並在反應器中持續 進行後續的作用程序； 添加作用菌體（2)，藻類的等分數量於反應器之 中並添加預定菌種，而菌體與藻類之間的比值係以1 : 3做為作用的量值比例，且環境溫度係設定為3 0至3 7度之間，水相反應，而該菌種係具有分解醣類並製造 出油脂之特性，其菌種係可使用酵母菌或絲狀真菌，而 使用的酵母菌分別係可為：彎隱球酵母（Cryptococcus albidus)、斯達氏油脂酵母（Lipomyces starkeyi 099132365 表單編號A0101 第5頁/共13頁 0992056705-0 201213546 )、出芽絲孢酵母（丁1*丄(：|1〇5口〇1'011卩11111113118)、產 油油脂酵母（Lipomyces lipofer)等，使用的絲狀真 菌則分別係可為：土曲霉（Aspergil lus ter-re128 ) 、暗頁枝孢霉（Cla-dosprium. fulvum)、花冠蟲霉 (Entomphl-hora coyonata)、梨形卷旋枝霉（ Helicostylum phrifome)、爪岐毛霉（Mucor javanicus)等’而該各菌種於反應器中並可與藻類進 行作用； 生物轉換作用（3) ’在預定菌體進入反應器與藻 類接觸後，該預定的菌體會分泌氣維水鱗酵素、澱粉分 .. .. 解酵素、脂肪合成酵素等，所述之該各酵素 ，例如：脂[0004] The effect is relatively long, and only the occurrence of variables in the implementation process can achieve the highest practical effect. More detailed explanation is that the more formalities, the more chances of failure (ie, variables) are added. In the present invention, after the preparation of the algae, the natural conversion between the algae and the organism of the microbial organism is directly produced to produce the crude oil, and about 20 to 50% of the algae are cellulose, algin and polysaccharide. Body and other substances, so the amount of crude oil in the material after conversion can be effectively higher than the prior art to increase the amount of oil, which improves the efficiency of the overall process, and the raw crude oil also provides the human demand The energy oil is used to achieve environmental benefits, energy conservation and other benefits, and in line with industrial use. [Embodiment] In order to more clearly express the technical gist of the present invention and to achieve the technical means to be used, a more detailed description will be given in the following with the accompanying drawings: The present invention relates to a microbial conversion of algal saccharides to obtain raw crude oil. The method, [refer to the first figure] mainly comprises: algae preparation (1), which is to carry out large-scale cultivation of predetermined algae, and is placed in the reactor in an appropriate amount during the implementation, and the algae used in the present invention The non-oily algae is mainly used, and the algae continues to carry out the subsequent action procedure in the reactor; the added bacteria (2), the aliquot of the algae is added to the reactor and the predetermined strain is added, and the bacteria are The ratio between the algae is a ratio of the ratio of 1:3, and the ambient temperature is set to between 30 and 37 degrees, and the aqueous phase reacts, and the strain has a decomposition sugar and produces a fat. The characteristics of the strain may be yeast or filamentous fungi, and the yeasts used may be: Cryptococcus albidus, S. cerevisiae (Lipomyces starkeyi 099132365) No. A0101 Page 5 of 13 0992056705-0 201213546 ), Trichosporon sp. (Ding 1*丄(:|1〇5〇1'011卩11111113118), Oil-producing oil yeast (Lipomyces lipofer), etc. The filamentous fungi can be: Aspergil lus ter-re128, Cla-dosprium. fulvum, Entomphl-hora coyonata, Trichosporon serrata ( Helicostylum phrifome), Mucor javanicus, etc. and the strains are in the reactor and can interact with the algae; bioconversion (3) 'after the predetermined cells enter the reactor and contact the algae, The predetermined bacteria will secrete gas-reducing water-scale enzymes, starch fractions, enzymes, fat-synthesizing enzymes, etc., and the various enzymes, such as lipids.
酸合成酶（fatty acid synthase)、乙稀輔酶A叛 化酶（acetyl CoA carboxylase)、固醇輔酶A去飽 和酶-1 (stearoyl CoA desaturase-1)以及與膽固 醇生合成相關酵素的戊二酸輔酶A合成酶（HMGC〇A ynthase)、戊二酸輔酶環原酶（hmgc〇a reductase) 等正向調控 ，以對該藻類體内的纖維素 、非澱 粉類多酷體、藻勝等物質進行物質轉換成醣體，醣體繼 以供菌體透過脂肪合成酵素等製造生質原油； 取得產物（4)，即係該被製造出的生質原油取出 ，而該生質原油則可再透過轉脂化成為生質柴油者。 本發明以微生物轉化藻類醣質取得生質原油之方法 ，〔請參閱第一圖〕簡單地說，就是將藻類製備（工） 後，將預定藻類適量地置入在反應器中，而假設在正常 生物反應器下進行運作，單次置入藻類的量為丄5一3 099132365 0 g/L，繼而在該反應器内添加作用菌體（2 ) 表單編號A0101 第6頁/共13頁 ，由於作 0992056705-0 201213546 用菌體添加量係與藻類之間比值呈1:3來執行，故相 對地，單次置入菌體的量為5 —i 0g/L，在此分別假設 藻類置入的量為1 5g/L、菌體置入的量為5g/L，讓該 菌體與藻類之間進行生物轉換作用（3)，該所述之轉 換作用係由於該菌體會分泌纖維水解酵素、澱粉分解酵 素等，以對該藻類體内的纖維素、非澱粉類多醣體、藻 膠等物質進行物質轉換成醣體，醣體繼以供菌體透過脂 肪合成酵素等製造生質原油，經此，該被製造出的生質 原油並可取出進行利用者》Fatty acid synthase, acetyl CoA carboxylase, stearoyl CoA desaturase-1, and glutarate coenzyme associated with cholesterol biosynthesis A synthetase (HMGC〇A ynthase), glutaric acid coenzyme loopase (hmgc〇a reductase) and other positive regulation, in order to carry out the cellulose, non-starch genus, algae and other substances in the algae The substance is converted into a saccharide body, which is followed by the production of raw crude oil by the bacteria through the fat synthesis enzyme; the product (4) is obtained, that is, the produced crude oil is taken out, and the raw crude oil is re-transmissible. Turn fat into biodiesel. The invention adopts a method for microbial conversion of algal saccharide to obtain raw crude oil, [refer to the first figure]. Simply put, after preparing the algae, the predetermined algae is placed in the reactor in an appropriate amount, and it is assumed that Working under a normal bioreactor, the amount of algae placed in a single time is 丄51-39913365 0 g/L, and then the cells are added to the reactor (2) Form No. A0101 Page 6 of 13 Since the ratio of the addition amount of the bacteria to the algae is 1:3, the amount of the single-incorporation of the bacteria is 5 - i 0g / L, and the algae are assumed here. The amount of the incorporation is 15 g/L, and the amount of the cells is 5 g/L, and the bacteria and the algae are subjected to bioconversion (3), and the conversion is due to the hydrolysis of the cells by the cells. Enzymes, amylolytic enzymes, etc., to convert substances such as cellulose, non-starch polysaccharides, algae and the like into sugar bodies in the algae, and the sugar bodies are used to produce raw crude oil through the fat synthesis enzymes. Through this, the produced crude oil can be taken out Conducting users
Ο 此外，因為藻類體内所含脂肪的前驅物約只有整體 的1 0%以下，故對於單次置入藻類的量為丄5g/L，以 及菌體置入的量為5 g/L來說，菌體實際上作用的僅有藻 類整體量的10% (含以下）’亦即，菌體實際上仍可 再對9 0% (含以上）的藻類脂肪的前驅物進行作用， 藉之，藻類置入的量可達1 5 0g/L，g當然不能一次就 將1 5 0 g/L的藻類量置入，必須分階段來置入以避免產 生受質抑制的現象，此可更徹底的讓|體與藻類之間達 到最佳的作甩效果： 甚者，當在使用的菌體其生物特性較為穩定的情況 下，可分別對藻類與菌體進行濃縮，〔請參閱第二圖〕 該係將藻類製備（1)後，將進行濃縮藻類，繼 而把預定藻類適量地置入在反應器中，由於藻類已經過 濃縮，故其置入的藻類量可達丄5 〇g/L ,繼而同樣濃縮 作用菌體（5),再對該反應器内添加作用菌體（ ，由於作用菌體添加量係與藻類之間比值呈1:3來執 099132365 行，故相對地，單次置入濃縮菌體的量為5 〇g/L， 表單編號A0101 第7頁/共13頁 讓該 0992056705-0 201213546 菌體與藻類之間進行生物轉換作用（3)，該所述之轉 換作用係由於該菌體會分泌纖維水解酵素、澱粉分解酵 素等，以對該藻類體内的纖維素、非澱粉類多醣體、藻 膠等物質進行物質轉換成醣體，醣體繼以供菌體透過脂 肪合成酵素等製造生質原油，經此，該被製造出的生質 原油並可取出進行利用，又，亦因藻類體内所含脂肪的 前驅物約只有整體的1 5 0%以下，故藻類置入的量可 再提高達到1 5 0 Og/L，其亦必須分階段來置入以避免 產生受質抑制的現象，藉此，以反應器單次作用的量而 言，相當可觀者。 由於在整體萃油過程中，談藻類不需透過培養基、 環境等因素來改變其藻類個體的生物組成，例如：藉由 減氮或透過培養基的方式來讓藻類體内所含的油脂增加 ，故其整體萃油流程至少減少了一至多道手續，此外， 在進行作用時係屬於生物體之間的自然轉換作用，而藻 類體内約有2 0〜5 0 %以上係為纖維素、藻膠以及多 醣體等非油物質，故在其物質經由轉換後的生質原油量 係可有效達到較高於以往技術來提高油脂量，故在整體 流程更有效提升了生產的整體效率，且同時也提供了人 類所需求的生質原油，此不僅符合環保概念，更可助於 延緩人類開採地球資源以同時有效延續地球與人類的生 存者。 綜上所述，當知本發明具有新穎性，且本發明未見 之於任何刊物，當符合專利法第2 1、2 2條之規定。 唯以上所述者，僅為本發明之一較佳實施例而已， 當不能以之限定本發明之範圍。即大凡依本發明申請專 099132365 表單編號A0101 第8頁/共13頁 0992056705-0 201213546 利範圍所作之均等變化與修飾，皆應仍屬本發明專利涵 蓋之範圍内。 【圖式簡單說明】 [0005] 第一圖：本發明藻類與菌體作用取得生質原油之流程示 意圖 第二圖：本發明另一藻類與菌體作用取得生質原油之流 程示意圖 【主要元件符號說明】 D [0006] 1 藻類製備 2 添加作用菌體 3 生物轉換作用 4 取得產物 5 濃縮藻類 6 濃縮作用菌體 Ο 099132365 表單編號A0101 第9頁/共13頁 0992056705-0Ο In addition, because the precursor of fat contained in algae is only about 10% of the whole, the amount of algae placed in a single time is 丄5g/L, and the amount of bacteria is 5 g/L. It is said that only 10% (including the following) of the total amount of algae actually acts on the bacteria', that is, the bacteria can actually act on the precursors of 90% (including above) algae fats. Algae can be placed in an amount of up to 150g/L. Of course, it is not possible to put 10.5 g/L of algae at a time. It must be placed in stages to avoid the occurrence of matrix inhibition. Thoroughly achieve the best effect between the body and the algae: In addition, when the biological characteristics of the cells used are relatively stable, the algae and the bacteria can be separately concentrated, [see second Fig.] After the algae is prepared (1), the algae will be concentrated, and then the predetermined algae will be placed in the reactor in an appropriate amount. Since the algae has been concentrated, the amount of algae placed can reach 〇5 〇g/ L, and then concentrate the bacteria (5), and then add the bacteria to the reactor (due to the action bacteria) The ratio between the body addition amount and the algae is 1:3 to hold the 099132365 line, so the amount of the single-stage concentrated bacteria is 5 〇g/L, Form No. A0101 Page 7 of 13 0992056705-0 201213546 Biotransformation between bacteria and algae (3), the conversion is due to the secretion of fibrinolytic enzymes, amylolytic enzymes, etc., to the cellulose in the algae, non- A substance such as a starch-based polysaccharide or an algal rubber is converted into a saccharide, and the saccharide is used to produce a raw crude oil through a fat synthesis enzyme, whereby the produced crude oil can be taken out and used. Moreover, because the precursor of fat contained in algae is only about 150% of the whole, the amount of algae can be increased to 150 Kg/L, which must be placed in stages to avoid The phenomenon of inhibition of the host is produced, whereby the amount of the single action of the reactor is considerable. Since the algae does not need to change the organism of the algae by the medium or the environment during the whole oil extraction process. Composition, for example: by subtraction Nitrogen or permeate the medium to increase the amount of oil contained in the algae, so the overall extraction process is reduced by at least one or more procedures. In addition, it acts as a natural conversion between organisms, while algae About 20 to 50% of the body is a non-oil substance such as cellulose, algin, and polysaccharide. Therefore, the amount of crude oil in the substance after conversion can be effectively higher than the prior art to improve the fat. Quantity, so the overall process is more effective to improve the overall efficiency of production, but also provides the raw crude oil required by humans, which not only conforms to the concept of environmental protection, but also helps to delay the exploitation of earth resources by humans to effectively continue the earth and humanity. Survivor. In summary, it is known that the present invention is novel, and the present invention is not found in any publication, and is in compliance with the provisions of Articles 21 and 22 of the Patent Law. The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto. That is, the application of the invention is based on the invention. 099132365 Form No. A0101 Page 8 of 13 0992056705-0 201213546 The equal changes and modifications made by the scope of the invention are still within the scope of the patent of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS [0005] First: Schematic diagram of the process of obtaining algin crude oil by the action of algae and bacteria in the present invention. Second diagram: Schematic diagram of the process of obtaining biomass crude oil by another algae and bacteria in the present invention [main components] Explanation of symbols] D [0006] 1 Preparation of algae 2 Addition of bacteria 3 Biotransformation 4 Obtaining product 5 Concentrated algae 6 Concentration of bacteria Ο 099132365 Form No. A0101 Page 9 of 13 0992056705-0