JPH0235074A - Hybrid chlorella cell produced from limnetic and marine chlorella cell and method for cell fusion thereof - Google Patents
Hybrid chlorella cell produced from limnetic and marine chlorella cell and method for cell fusion thereofInfo
- Publication number
- JPH0235074A JPH0235074A JP63184623A JP18462388A JPH0235074A JP H0235074 A JPH0235074 A JP H0235074A JP 63184623 A JP63184623 A JP 63184623A JP 18462388 A JP18462388 A JP 18462388A JP H0235074 A JPH0235074 A JP H0235074A
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- Japan
- Prior art keywords
- cells
- chlorella
- cell
- freshwater
- chlorella cells
- Prior art date
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Links
- 241000195649 Chlorella <Chlorellales> Species 0.000 title claims abstract description 100
- 230000007910 cell fusion Effects 0.000 title claims abstract description 16
- 238000000034 method Methods 0.000 title claims description 8
- 210000004027 cell Anatomy 0.000 claims abstract description 111
- 210000002421 cell wall Anatomy 0.000 claims abstract description 15
- 210000001938 protoplast Anatomy 0.000 claims abstract description 13
- 230000001172 regenerating effect Effects 0.000 claims abstract description 3
- 239000013505 freshwater Substances 0.000 claims description 37
- 239000013535 sea water Substances 0.000 claims description 21
- 238000003163 cell fusion method Methods 0.000 claims description 7
- 230000004927 fusion Effects 0.000 claims description 4
- 235000020673 eicosapentaenoic acid Nutrition 0.000 abstract description 10
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 abstract description 9
- 229960005135 eicosapentaenoic acid Drugs 0.000 abstract description 9
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 abstract description 9
- 238000006243 chemical reaction Methods 0.000 abstract 1
- 239000003795 chemical substances by application Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 229930195725 Mannitol Natural products 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 230000008929 regeneration Effects 0.000 description 3
- 238000011069 regeneration method Methods 0.000 description 3
- 239000000600 sorbitol Substances 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 229960004793 sucrose Drugs 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 229930002875 chlorophyll Natural products 0.000 description 1
- 235000019804 chlorophyll Nutrition 0.000 description 1
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000013367 dietary fats Nutrition 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
Landscapes
- Edible Seaweed (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
（イ）産業上の利用分野
この発明は、淡水産クロレラ細胞と海水産クロレラ細胞
両種の雑種クロレラ細胞、及びその細胞融合法に関する
ものである。DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application The present invention relates to hybrid chlorella cells of both freshwater and seawater chlorella cells, and a cell fusion method thereof.
（１１）従来の技術
クロレラは単細胞緑藻てあり、・淡水産クロレラ細胞と
海水産クロレラ細胞に大別分類することかてきる。(11) Conventional technology Chlorella is a unicellular green alga, and can be broadly classified into freshwater chlorella cells and saltwater chlorella cells.
−・般にクロレラと云う場合、淡水産クロレラ細胞を指
していることか多い。クロロフィルを多く含み、タンパ
ク質含有料も４０〜５０％と多く、増殖速度か早いため
採算に合い易く大贋培養される機会も多いためである。-Generally, when we say chlorella, we are often referring to freshwater chlorella cells. This is because it contains a lot of chlorophyll, has a high protein content of 40 to 50%, and has a fast growth rate, making it profitable and often being cultivated in large quantities.
その栄養価の高さと増殖速度の早さから、宇宙食として
実用の方向へ研究か進められていることは周知のところ
である。It is well known that due to its high nutritional value and rapid growth rate, research is underway to put it into practical use as space food.
一方の海水産クロレラ細胞であるか、一般にはあまり知
られていない。本願発明の出願人は昭和５８年より海水
産クロレラ細胞の大量培養研究開発に着手し、翌年には
海水産クロレラ細胞の優良株の分離に成功した。海水産
クロレラＭｕ胞の特色としては脂質含有か多く、特に人
体有用゛′Ｘ：養脂質であるＥＰＡ （エイコサペンタ
エン酸）かつ富である。On the other hand, it is not well known to the general public that it is a seawater chlorella cell. The applicant of the present invention began research and development on mass culture of marine chlorella cells in 1981, and succeeded in isolating an excellent strain of marine chlorella cells the following year. Seawater Chlorella Mu cells are characterized by a high content of lipids, and are especially rich in EPA (eicosapentaenoic acid), which is useful for the human body.
（ハ）発１９１か解決しようとする課題＋ｉｉｊ記した
ように、淡水産クロレラ細胞には含まれない人体イｊ用
栄養脂質であるＥＰＡを多く含んた海水産クロレラ細胞
てはあるか、淡水産クロレラ細胞に比較して酢酸・ット
ウ糖等、炭素源を資化する走力か低く、従って増殖速度
も極めて遅いのである。海水産クロレラ細胞を大漬培養
するためには淡水産クロレラ細胞の何倍ものコストか掛
かり、低廉に供給することは現１１４点ては不可俺な状
態である。(c) Problems to be solved +iii As mentioned above, are there any seawater chlorella cells that contain a large amount of EPA, a nutritional lipid for the human body that is not contained in freshwater chlorella cells? Compared to Chlorella cells, they have a lower ability to assimilate carbon sources such as acetic acid and cane sugar, and therefore their growth rate is extremely slow. It costs many times more to culture saltwater chlorella cells than freshwater chlorella cells, and it is currently impossible to supply them at a low cost.
たとえば、淡水産クロレラ細胞の増殖速度は２０〜３０
ｇ／ｒｎ’ｌｌ （藻体生産性）と！ｉｊ （、これに
対して、海水産クロレラ細胞の増殖速度は２〜３ｇ／は
口（藻体生産性）と極めて遅い。For example, the growth rate of freshwater Chlorella cells is 20-30
g/rn'll (algal productivity)! ij (On the other hand, the growth rate of seawater Chlorella cells is extremely slow at 2 to 3 g/mouth (algal body productivity).
この発１」は、増殖速度の早い淡水産クロレラ細１泡と
組Ｉ＆成分に多くのＥＰＡを含む海水産クロレラ細胞と
の両種の特色を保有するところの雑種クロレラ細胞と、
その細胞融合法を提供することを［１的とするものであ
る。This development 1 is a hybrid chlorella cell that possesses the characteristics of both freshwater chlorella cells, which have a fast multiplication rate, and seawater chlorella cells, which contain a large amount of EPA in group I and components.
Our first objective is to provide a method for cell fusion.
（ニ）課題を解決するだめの手段
この発明は、淡水ｌｒクロレラ細胞と７に水産クロレラ
細胞の両種を細胞融合して作出されることを特徴とする
雑種クロレラ細胞に関するものである。(D) Means for Solving the Problems The present invention relates to a hybrid chlorella cell which is produced by cell fusion of freshwater lr chlorella cells and aquatic chlorella cells.
また、この発明は、原形質体を残して細胞壁だけを溶解
させる淡水産クロレラ細胞のプロトブラスト化、同しく
原形質体を残して細胞壁だけを溶解させる海水産クロレ
ラ細ＩＤのプロトブラスト化、つぎに淡水産クロレラプ
ロトプラス１〜細胞と海水産クロレラプロトブラスト細
胞を細胞融合させ、そして淡水産クロレラブロトフラス
ト細胞と海水産クロレラプロトブラスト細胞との融合細
胞に細胞ｊｔｔ ？再生させるという各Ｉ：程から成る
ことを特徴とする。淡水産クロレラ細胞と海水産クロレ
ラ細胞の細胞融合法に関するらのである。In addition, this invention involves protoblastization of freshwater Chlorella cells, which dissolves only the cell wall while leaving the protoplasts; Freshwater Chlorella protoplus 1 ~ cells and marine Chlorella protoblast cells are cell fused, and the cells jtt? It is characterized by consisting of each I: to regenerate. This paper describes a cell fusion method between freshwater Chlorella cells and saltwater Chlorella cells.
（・ト）作用
１；述のような、淡水産クロレラ細１抱１ と海水産ク
ロレラ細胞４の両種を細胞融合した雑種クロレラ細胞１
１によれば、ＥＰＡは海水産クロレラ、細胞４と同様な
含＋、）を有し、しかも、淡水産クロレラ細胞１並みの
増殖速度を持っており、大清培養がＩｖＴｆ七である。(G) Effect 1; Hybrid chlorella cell 1, which is a cell fusion of both freshwater chlorella cells 1 and seawater chlorella cells 4, as described above.
According to No. 1, EPA has the same content as seawater Chlorella cell 4, has a growth rate comparable to that of freshwater Chlorella cell 1, and has an IvTf of 7 when cultured in large serum.
また、淡水産クロレラ細胞１と海水産クロレラ細胞４の
細胞融合法であるか、まず、細胞融合の邪魔になるト１
４細胞の外側を覆う固い細胞壁３を溶解してしまって、
両細胞の原形質体２のみをのこす（プロドブロスト化）
細胞壁３を溶解してしまった淡水イクロレラグロトプラ
ス１〜細胞６とＷ６水産クロレラプロトブラスト細胞７
は容易に細胞融合させることかてきる。In addition, whether it is a cell fusion method of freshwater Chlorella cells 1 and seawater Chlorella cells 4, first of all, there are
4 dissolves the hard cell wall 3 that covers the outside of the cell,
Leaving only the protoplasts 2 of both cells (prodoblast formation) Freshwater Chlorella glotoplasts 1 to 6 with cell walls 3 dissolved and W6 aquatic Chlorella protoblast cells 7
can easily cause cell fusion.
つぎに、淡水産クロレラプロトブラスト細胞６と海水産
クロレラプロトプラス１〜細胞７との融合細胞の外側を
覆う細胞壁３を再生させることで、雑種クロレラ細胞１
！か作出される。Next, by regenerating the cell wall 3 that covers the outside of the fused cell of the freshwater Chlorella protoblast cells 6 and the seawater Chlorella protoplus cells 1 to 7, the hybrid Chlorella cells 1
! Or created.
い）実施例
この発明に係る淡水産クロレラ細胞と１１４水産クロレ
ラ細胞から作出される雑種クロレラ細胞及びその細胞融
合法の実施例を添付図面に基づいて説明するか、これら
の実施例のみに限定されるものでないことは云うまても
ない。b) Examples Examples of the hybrid chlorella cells produced from freshwater chlorella cells and 114 aquatic chlorella cells and the cell fusion method thereof according to the present invention will be explained based on the attached drawings, or will be limited only to these examples. Needless to say, this is not the case.
第１図は、淡水産クロレラ細胞ＩとｄＩｉ水産クロレラ
細胞４をあられず。Figure 1 shows freshwater Chlorella cells I and dIi Aquatic Chlorella cells 4.
第１ ［Ａ中１は原形質体２の外側を固い細胞１Ｖ３に
覆われた淡水産クロレラ細胞である。1st [1 in A is a freshwater Chlorella cell in which the outside of the protoplast 2 is covered with hard cells 1V3.
第１１ｇ ｉｔ ４は原形質体２の外側を固い細胞壁３
に覆われた淡水産クロレラ細胞である。The 11th g it 4 has a hard cell wall 3 on the outside of the protoplast 2.
These are freshwater chlorella cells covered with
第２図は、細胞壁溶解作用を持った酵素材５によって、
原形質体２・２のみな残しプロトプラス１−化した淡水
産クロレラプロ１〜ブラスト細胞６と海水産クロレラプ
ロ１−プラスＩ−ｌｉ８胞７をあられず。FIG. 2 shows that the enzyme material 5 having a cell wall dissolving action is used to
Freshwater Chlorella pro1-blast cells 6 and seawater Chlorella pro1-plus I-li8 cells 7, which were converted to protoplast 1- with only protoplasts 2 and 2 left, were found.
以下はプロトブラスト化条件の実験結果である。Below are the experimental results of protoblastization conditions.
細胞濃度は、淡水産クロレラ細胞ｌか１．０〜２ 、
ＯＸ １０ ’ｌＣｅ１ｌｓ／−ｌ 、海水産クロレラ
細胞４か０．５〜１．０Ｘ１０°Ｃｅ１ｌｓ／ｍｌに良
好な結果か得られた。The cell concentration is 1.0-2 liters of freshwater chlorella cells,
Good results were obtained with 0.5 to 1.0 x 10°Ce1ls/ml of seawater Chlorella cells.
酵素剤５としてセルラーゼとペクチナーゼを用いた。各
々４．０％を一応の基準とした。Cellulase and pectinase were used as enzyme agent 5. The standard was set at 4.0% for each.
プロトブラスト安定剤として０．４鵠〜０゜６Ｍソルビ
トール／マンニトール（１： ｌ）溶液に良好な結果か
得られた。Good results have been obtained with 0.4 to 0.6M sorbitol/mannitol (1:1) solution as a protoblast stabilizer.
種々のＰｌ＋のリン酸バッファーを使用して実験したと
ころ、プロトブラスト化のための四（は５６〜６．４に
良好な結果が得られた。When experimenting using various Pl+ phosphate buffers, good results were obtained for protoblastization from 56 to 6.4.
プロトブラスト化温度としては２５℃〜３０°Ｃに良好
な結果か得られた。Good results were obtained at a protoblasting temperature of 25°C to 30°C.
プロｌルブラスト化のための酵素反応時間としては１２
時間〜１８時間に良好な結果か得られた。The enzyme reaction time for prolublastization is 12
Good results were obtained between hours and 18 hours.
第３ Ｆ／Ｉ （＾）（Ｉｔ）（Ｇ）は融合剤８の働き
によって淡水産クロレラプロトブラスト細胞６とＷ＋ｊ
水産クロりラプロトブラスト細１泡７の細胞融合か進行
する過程をあられし、（＾）は互いに引合う状態、（ロ
）は細胞融合の進行途中の状態、（ｃ）は両種の原形質
体２・２か完全に融合して一つの融合細胞９に成った状
態をあられす。3rd F/I (^) (It) (G) is combined with freshwater Chlorella protoblast cells 6 and W+j by the action of fusion agent 8.
The process of cell fusion in marine black laprotoblast fine 1 bubble 7 is shown. (^) is a state where they are attracted to each other, (b) is a state in the process of cell fusion, and (c) is the origin of both species. Hail, the plasma bodies 2.2 have completely fused into one fused cell 9.
以下は細胞融合条件の実験結果である。Below are the experimental results of cell fusion conditions.
プロトブラスト細胞濃度は１．０ｘｌｏ’〜１ 、 Ｏ
Ｘ Ｉ Ｏ’Ｃｅ１ｌｓ／ｍｌの範囲て良好な結果を得
られた。Protoblast cell concentration ranges from 1.0xlo' to 1,0
Good results were obtained within the range of X I O'Ce1ls/ml.
融合剤８として１００ ｍ１ｌｌｔｌｌ化カルシウム、
０６１ＩＩソルビトール／マンニトール（１： ｌ）及
び３０％ポリエチレンクリコール６０００の緩衝液を使
用した。100 ml calcium chloride as fusing agent 8;
A buffer of 061II sorbitol/mannitol (1:1) and 30% polyethylene glycol 6000 was used.
１ン１１は１００ｍＭ塩化カルシウムをＩＮ塩酸てｐＨ
８，０〜８．５に調整した。1-11 is 100mM calcium chloride in hydrochloric acid pH
It was adjusted to 8.0 to 8.5.
細胞融合は２５℃〜３０℃の温度て起ることか確認され
た。It was confirmed that cell fusion occurred at a temperature of 25°C to 30°C.
細胞融合は３０分〜１時間て行われること力）確認され
た。It was confirmed that cell fusion was carried out for 30 minutes to 1 hour.
第４１Ａは、プロトブラスト再生培地１０に於て新たな
細胞壁３か１１生じ、雑種クロレラ細胞１１か作出され
た状態をあられしている。No. 41A shows a state in which new cell walls 3 or 11 are generated in the protoblast regeneration medium 10 and hybrid Chlorella cells 11 are produced.
本願出願人か実験に用いたプロトブラスト１り生培地Ｉ
Ｄは０．６１４ソルビトール／マンニトール（１：１）
、１０％シュクロース、５％寒天を含んており、ここに
融合細胞９を重層して蛍光灯照明ド室温て２週間程培養
して行なったまた、実験に用いたプロトブラスト再生培
地ＩＯに含まれる微量成分を説ＩＪ１すれば、０．５％
尿素、０．１％硫酸マグネシクム、０．０５％リン酸−
カリウム、ｏ、ｏｏｏｓ％硫酸第一鉄である。Protoblast 1 live medium I used in the experiment by the applicant
D is 0.614 sorbitol/mannitol (1:1)
, 10% sucrose, and 5% agar, and the fused cells 9 were layered thereon and cultured at room temperature under fluorescent lighting for about 2 weeks. According to IJ1, 0.5% of trace components
Urea, 0.1% magnesium sulfate, 0.05% phosphoric acid -
Potassium, o,oos% ferrous sulfate.
以１−か淡水産クロレラ細胞ｌと１ｍ水産クロレラ細胞
４を細胞融合する全工程である。The following is the entire process of cell fusion of 1-1 freshwater chlorella cells 1 and 1 m aquatic chlorella cells 4.
淡水産クロレラ細胞１と海水産クロレラ細胞４から作出
された雑種クロレラ細胞１１は１両種の特性を兼備して
いる。即ち、ＥＰＡの含有ｒＩ）の多さと増殖速度の１
Ｐさである。Hybrid chlorella cells 11 produced from freshwater chlorella cells 1 and seawater chlorella cells 4 have the characteristics of both species. That is, the amount of rI contained in EPA and the growth rate are 1
It's P.
（ト）発明の効果
今まて説明して来たように、従来ＥＰＡ含有場の多い海
水産クロレラ細胞の増殖速度は淡水産クロレラ細胞に比
して極めて遅いという欠点かあった。(G) Effects of the Invention As explained above, conventionally, the growth rate of saltwater Chlorella cells containing many EPA-containing sites was extremely slow compared to freshwater Chlorella cells.
しかし、この発明に係る淡水産クロレラ細胞と海水産ク
ロレラ細胞の細胞融合υ、によれば淡水産クロレラ細胞
の持つ増殖速度の早さと１１η水産クロレラ細胞の持つ
ＥＰＡを多く含有するという両特質を兼備えた、雑種ク
ロレラ細胞を作出することかてきる。However, according to the cell fusion υ of freshwater chlorella cells and seawater chlorella cells according to the present invention, it has both the fast proliferation rate of freshwater chlorella cells and the high EPA content of 11η aquatic chlorella cells. It is possible to create hybrid chlorella cells with the following features.
°Ｉ覧実、雑種クロレラ細胞のＥＰ’Ａ含有丑含有水産
クロレラ細胞と同様に４〜７％であることか確認されて
いる。In fact, it has been confirmed that the EP'A-containing hybrid chlorella cells have a content of 4 to 7%, similar to the EP'A-containing aquatic chlorella cells.
以」；のように、この発明に係る淡水産クロレラ細胞と
海水産クロレラ細胞から作出される雑種クロレラ細胞、
及びその細胞融合法の産業−１−の利用価値は極めて高
いものと云わねばならないHybrid chlorella cells produced from freshwater chlorella cells and seawater chlorella cells according to the present invention,
It must be said that the utility value of the cell fusion method in industry-1- is extremely high.
添付図面はこの発明に係る淡水産クロレラ細胞と海水産
クロレラ細胞から作出される雑種クロレラ細胞、及びそ
の細胞融合法の実施例をあられし、第１図は淡水産クロ
レラ細胞とｄ６水産クロレラ細胞の参考図、第２図はプ
ロトブラスト化の参考図、第３図は細胞融合の参考図、
第４図は細胞壁再生の参考図を夫々あられしている。
第１
図
第３図
第２図
図中の主な符号
（１）・・・淡水産クロレラ細胞、（２）・・・原形質
体、（３）・・・細胞壁、 （４）・・・海水産クロレ
ラ細胞。
（５）・・・酵素剤、
（６）・・・淡水産クロレラプロトブラスト細胞、（７
）・・・海水産クロレラプロドブロスト細胞、（８）・
・・融合剤、 （９）・・・融合細胞、（１０）・・
・プロトブラスト再生培地、（１１）・・・雑種クロレ
ラ細胞。
第４図
「統補正ｉ８（方式）
％式％
：１
２、発明の名称
３、補１Ｆをする者
・ＩＳ件との関係The attached drawings show examples of hybrid chlorella cells produced from freshwater chlorella cells and seawater chlorella cells, and the cell fusion method thereof, according to the present invention. Reference figure, Figure 2 is a reference figure for protoblastization, Figure 3 is a reference figure for cell fusion,
Figure 4 shows reference diagrams for cell wall regeneration. Main symbols in Figure 1 Figure 3 Figure 2 (1)...Freshwater Chlorella cells, (2)...Protoplasts, (3)...Cell walls, (4)... Marine chlorella cells. (5) Enzyme agent, (6) Freshwater Chlorella protoblast cells, (7
)...Marine Chlorella prodobrost cells, (8)
...Fusion agent, (9)...Fusion cell, (10)...
- Protoblast regeneration medium, (11)...Hybrid Chlorella cells. Figure 4 “Stationary amendment i8 (method) % formula %: 1 2. Title of the invention 3, person making supplement 1F/relationship with IS matter
Claims (1)
細胞融合して作出されることを特徴とする雑種クロレラ
細胞 ２、以下の各工程から成ることを特徴とする、淡水産ク
ロレラ細胞と海水産クロレラ細胞の細胞融合法 （ａ）原形質体を残して細胞壁だけを溶解させる淡水産
クロレラ細胞のプロトブラスト化工程 （ｂ）原形質体を残して細胞壁だけを溶解させる海水産
クロレラ細胞のプロトブラスト化工程 （ｃ）淡水産クロレラプロトブラスト細胞と海水産クロ
レラプロトブラスト細胞を細胞融合させる工程 （ｄ）淡水産クロレラプロトブラスト細胞と海水産クロ
レラプロトブラスト細胞との融合細胞に細胞壁を再生さ
せる工程[Claims] 1. A hybrid chlorella cell characterized by being produced by cell fusion of both freshwater chlorella cells and seawater chlorella cells 2. A hybrid chlorella cell characterized by comprising the following steps: Cell fusion method of freshwater Chlorella cells and seawater Chlorella cells (a) Protoblastization process of freshwater Chlorella cells, leaving the protoplasts and dissolving only the cell walls (b) Dissolving only the cell walls, leaving the protoplasts Step of protoblastization of seawater chlorella cells (c) Step of cell fusion of freshwater chlorella protoblast cells and seawater chlorella protoblast cells (d) Fusion cell of freshwater chlorella protoblast cells and seawater chlorella protoblast cells The process of regenerating cell walls in
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63184623A JPH0235074A (en) | 1988-07-26 | 1988-07-26 | Hybrid chlorella cell produced from limnetic and marine chlorella cell and method for cell fusion thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63184623A JPH0235074A (en) | 1988-07-26 | 1988-07-26 | Hybrid chlorella cell produced from limnetic and marine chlorella cell and method for cell fusion thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0235074A true JPH0235074A (en) | 1990-02-05 |
Family
ID=16156474
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63184623A Pending JPH0235074A (en) | 1988-07-26 | 1988-07-26 | Hybrid chlorella cell produced from limnetic and marine chlorella cell and method for cell fusion thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0235074A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5962121A (en) * | 1996-12-12 | 1999-10-05 | 3M Innovative Properties Company | Retroreflective sheet comprising microspheres, the diameter and refractive index of which being specifically related to the refractive indices of layers directly in contact therewith |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6232877A (en) * | 1985-08-06 | 1987-02-12 | Nisshin Oil Mills Ltd:The | Novel chlorella |
-
1988
- 1988-07-26 JP JP63184623A patent/JPH0235074A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6232877A (en) * | 1985-08-06 | 1987-02-12 | Nisshin Oil Mills Ltd:The | Novel chlorella |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5962121A (en) * | 1996-12-12 | 1999-10-05 | 3M Innovative Properties Company | Retroreflective sheet comprising microspheres, the diameter and refractive index of which being specifically related to the refractive indices of layers directly in contact therewith |
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