CN102286381A

CN102286381A - Method for breaking microalgal cell walls

Info

Publication number: CN102286381A
Application number: CN2010102027049A
Authority: CN
Inventors: 褚俊杰; 李顺来; 谢晓惠; 郑嘉惠
Original assignee: BIOLOGICAL RESOURCE Co Ltd
Current assignee: BIOLOGICAL RESOURCE Co Ltd
Priority date: 2010-06-18
Filing date: 2010-06-18
Publication date: 2011-12-21

Abstract

The invention discloses a method for breaking microalgal cell walls. The method mainly includes bacteria culture, microalgae addition, hydrolytic enzyme release, cell wall hydrolyzation and microalgal broth extraction, wherein bacteria culture is to culture a predetermined number of bacteria in culture solution in a predetermined space and add microalgae in the culture space in coordination with the number of the bacteria, microalgae addition is to add microalgae to be decomposed into the bacterial culture space, so that the bacteria can release hydrolytic enzyme, the hydrolytic enzyme can hydrolyze the cell walls of the microalgae, the cell walls are converted into saccharide after being decomposed by the hydrolytic enzyme, moreover, the cell walls of the microalgae can gradually produce holes due to decomposition, the oil in the microalgae can be floated to the surface of the culture solution culturing the bacteria when the aperture of the hole is larger than the volume of the oil, and the broth remaining in the microalgae can be extracted and classified for use after the cell walls are completely decomposed.

Description

The method of microalgae cell walls broken wall

Technical field

The present invention relates to a kind of method of microalgae cell walls broken wall, mainly relate to a biotechnology at little algae, algae, this technology of mat is beneficial to extract the content of this little algae, algae.

Background technology

Along with global economy development and population increase fast, human for the also increase thereupon of greasy demand, and present greasy source is mainly still based on plant, animal tallow, but because demand is very big, at present plant, animal tallow can't satisfy human wants, so prior art is directed to and obtains grease in the algae gradually.

Because the oleaginousness of most algae is high, is example with little algae, little algae has been rich in protein, fat and carbohydrate, and some algae is rich trace elements and mineral substance more, that is little algae has photosynthetic organs such as chlorophyll, can effectively utilize sun power by photosynthesis with H ₂O, CO ₂Be converted into organic compound with inorganic salt, therefore can absorb CO by little algae ₂Greenhouse effect ease up, and the modes of reproduction of little algae is based on the binary fission formula, cell cycle is shorter, also be easy to carry out large scale culturing, but because the suitable tool resistance of cell walls of little algae and hard and degradation resistant, and cell walls is made up of Mierocrystalline cellulose, and Mierocrystalline cellulose is unfavorable for being decomposed in vivo by human body, animal, then reduce the absorbed efficient of nutrition content that is rich in little algae, obtain the technology that the interior stoste of little algae just becomes quite worth research so how to decompose, destroy cell walls.

Learn at present, the method of fracturing cell walls is rough classifies mechanical process, physics method and chemistry and biochemical process, its mechanical process mainly uses mechanical action to destroy cell walls by the mode of grinding or cut, this method is useful in animal tissues, be unsuitable at microorganism, and easily destroy intracellular original liquid component indirectly; Physical laws uses various physical factors to destroy cell walls, it can be classified approximately methods such as freezing molten method, ultrasonication repeatedly, freeze molten method repeatedly by repeatedly the freezing room temperature that enters again of cell being dissolved, through making for several times the cell wall structure fragmentation repeatedly, this method is suitable for being used in histocyte, relatively poor for the microorganism cells effect, and the ultrasonic wave of ultrasonication rule by certain power make cell walls sharply concussion break, this method is simple to operate, but easily makes certain part susceptibility active substance sex change inactivation; Chemistry divides again with the biological chemistry rule the molten method of enzyme, methods such as chemosmosis method, the molten method of enzyme is decomposed cell walls by various lytic enzymes, this method causes product inhibition easily, and then it is low to influence the intracellular matter release rate, and lyase cost height, so be unsuitable for extensive utilization, then must select different lyases at different bacterial classifications, this has also limited the use range of this method relatively, and the permeability that the chemosmosis rule is to use organic solvent to change cell walls infiltrates content, though this method is comparatively gentle, but because the time is long, reduced efficient relatively, so no matter above-mentioned or additive method all has suitable improvement or progressive space at present.

Because above-mentioned disappearance drawback, the inventor thinks that it has necessity of demanding urgently correcting, then be engaged in the many years of experience that related products manufactures and designs with it, and the well-designed theory of always adhering to, studied at above bad place, through after constantly making great efforts, be the method for releasing microalgae cell walls broken wall of the present invention eventually, with the effect of better biotechnology lift technique.

Summary of the invention

Main purpose of the present invention is, a kind of method that can decompose the alga cells wall efficiently and not destroy entocyte is provided.

For reaching aforesaid purpose, the method of microalgae cell walls broken wall of the present invention is to come the hydrolysis cell wall structure by the cellulose hydrolysis ferment, described cellulose hydrolysis ferment can be from fungi, in bacterium or the animal, because secreted cellulose hydrolysis ferment kind and the output of fungi is maximum, so the present invention is the main thalline of implementing with the fungi, and method of the present invention mainly comprises yeast culture, add little algae, disengage hydrolytic enzyme, steps such as hydrolysis cell walls and the little algae stoste of taking-up, yeast culture with an amount of yeast culture in a predetermined space, then little algae to be decomposed is added in the space of this cultivation thalline, and little algae quantity of adding cooperates the quantity of thalline, to make thalline can in effective time limit, disengage the cell walls that hydrolytic enzyme decomposes little algae, and after the little algae sum that adds has all decomposed, still the thalline that leaves some amount is for sustainable cultivation of thalline and breeding voluntarily, and after the little algae sum that adds all decomposed, can take out the grease in little algae, contents such as protein, and simultaneously sustainablely add an amount of little algae again and come to come the hydrolysis cell walls for the cellulose hydrolysis ferment that thalline disengages, so that reach a circulation process, this compares old wall breaking technology, method of the present invention not only can be come the hydrolysis cell walls by relatively mild mode, so that little algae content is not destroyed or runs off, and then allow the interior content of little algae be absorbed fully or to utilize, in addition, owing to used sustainable round-robin operating process, so can save the step of continuous replacement flow process on the one hand, more can effectively promote the decomposition efficiency of cell walls on the other hand.

Description of drawings

Fig. 1: the schematic flow sheet of wall-breaking method of the present invention.

Embodiment

The method of the relevant a kind of microalgae cell walls broken wall of the present invention, (seeing also Fig. 1) its method mainly comprise yeast culture 1, add little algae 2, disengage hydrolytic enzyme 3, hydrolysis cell walls 4 with take out little algae stoste 5 and constitute; Wherein:

Yeast culture 1, but the kind of thalline is the microorganism based on the plain hydrolytic enzyme of eccrine fiber, for example: fungi, bacterium, radioactive rays bacterium etc., the thalline of this stage with a pre-determined quantity is incubated in the predetermined space by nutrient solution, make the thalline breeding of can constantly in this space, deriving, and cooperate the quantity of thalline in the space of cultivating, to add little algae then;

Add little algae 2, mainly little algae to be decomposed is added in the space of cultivating thalline, and the number of little algae quantity of being added is to cooperate thalline, make in the scope that the actual quantity of thalline can decompose the cell walls of little algae fully determine, so the quantity that little algae is added is that the thalline sum can decompose the cell walls of little algae in the numerical value fully, and after adding little algae, thalline just can disengage hydrolytic enzyme;

Disengage hydrolytic enzyme 3, after in the space of yeast culture, adding little algae, originally exist the thalline this space in just to disengage hydrolytic enzyme voluntarily, described hydrolytic enzyme is a Mierocrystalline cellulose amylase, this Mierocrystalline cellulose amylase cell walls that then can be hydrolyzed to little algae of interpolation;

Hydrolysis cell walls 4 mainly pass through the secreted hydrolytic enzyme of thalline with the cell walls decomposition and inversion saccharogenesis body on little algae, and decomposition course mainly decomposites hole gradually on cell walls, and along with timeliness increases, it is big that the aperture of hole also becomes gradually, continues it, just can take out little algae stoste by this hole;

Take out little algae stoste 5, because the microalgae cell wall comes decomposition and inversion to become carbohydrate by the secreted hydrolytic enzyme of thalline, in decomposition course, this microalgae cell wall can form hole gradually because of decomposition, grease in little algae then can be in the aperture of hole emersion is to the nutrient solution of yeast culture during greater than the grease volume, the stoste that remains in little algae then can be removed after cell walls decomposes fully and the use of classifying.

When using the method enforcement of microalgae cell walls broken wall, see also Fig. 1, at first must insert the nutrient solution of thalline in a predetermined space, and control suitable growing environment to carry out yeast culture 1, suppose that employed thalline is a fungi, fungi can constantly derive to breed in this culture space and accelerate, in fungi reaches this culture space can hold a preset value of fungi quantity the time, just can in this space, add little algae (step 2), and the quantity of adding can allow fungi still can maintain the preset value of fungi quantity after fully little algae being decomposed, because fungi can be disengaged hydrolytic enzyme (step 3) at the contact exotic, and described hydrolytic enzyme is the cellulose hydrolysis ferment, so the cell walls of little algae of adding can be decomposed, when little algae (step 4) during by ferment hydrolysis cell walls, on cell walls, can have the hole that differs in the aperture because of the degree of decomposing gradually, through this, because the grease proportion in little algae is less, so can be in the aperture of hole during greater than the grease volume, the first emersion of grease meeting is to nutrient solution, then by the color observation in the nutrient solution, whether the cell walls that can understand little algae is decomposed fully, after the cell walls of little algae is decomposed fully, the carbohydrate that stoste in little algae and cell walls are changed into after hydrolytic enzyme decomposes can filter the back and take out in this space, allow fungi after fully little algae being decomposed, still maintain the preset value of fungi quantity owing to add little algae quantity of little algae and be, so (sustainablely after the step 5) add little algae again to reach a complete circulation process in the little algae stoste of taking-up.In addition, the grease in this little algae is convertible into biofuel, and the carbohydrate that other guide thing and cell walls hydrolysis change into then can be as protective foods or feed, and the residual thing of remaining algae then can use as compost, uses little algae content is intactly used.

The method of microalgae cell walls broken wall of the present invention, its advantage is, because the cell walls of little algae decomposes by the secreted hydrolytic enzyme that goes out of thalline and changes into carbohydrate, so after cell walls is decomposed, protective foods that this little algae is made or feed can be beneficial to and be absorbed, while is the little algae of tool fishy smell originally not, and in hydrolytic process, this mushroom still can be derived simultaneously to breed in culture space and be accelerated, so under can little algae quantity and the two situation about cooperatively interacting of thalline quantity in the yeast culture when adding little algae, after this microalgae cell wall is decomposed by the secreted hydrolytic enzyme of mushroom fully, mushroom can maintain in the preset value, through this, can be constantly in the space of yeast culture 1, add little algae, and then reach a circulation process efficiently, by this, can effectively stablize on the one hand the efficient of the little algae stoste of taking-up, can save on the other hand at thalline quantity not sufficient and the program that must prepare again continues to make method of the present invention reach suitable progressive effect in culture space.

The above only is a preferred embodiment of the present invention, can not with qualification scope of the present invention.Promptly the equalization of doing according to claim of the present invention generally changes and modifies, and all should belong in the scope that the present invention contains.

Claims

1. the method for a microalgae cell walls broken wall, this method mainly comprise yeast culture, add little algae, disengage hydrolytic enzyme, the hydrolysis cell walls with take out little algae stoste step; Wherein:

Yeast culture, but thalline is to use the microorganism of the plain hydrolytic enzyme of eccrine fiber, this stage with the yeast culture of a pre-determined quantity in predetermined space the breeding so that thalline can constantly be derived in this space, and cooperate the quantity of thalline in the space of cultivating, to add little algae then;

Adding little algae, mainly be that little algae to be decomposed is added in the space of cultivating thalline, and after adding little algae, thalline just can disengage hydrolytic enzyme;

Disengage hydrolytic enzyme, in the space of yeast culture, add little algae after, originally exist the thalline this space in just to disengage hydrolytic enzyme voluntarily, this hydrolytic enzyme cell walls that then can be hydrolyzed to little algae of interpolation;

The hydrolysis cell walls mainly passes through the secreted hydrolytic enzyme of thalline with the cell walls decomposition and inversion saccharogenesis body on little algae, after cell walls is decomposed, just can take out little algae stoste then;

Take out little algae stoste, cell walls is by in the thalline decomposition course, this microalgae cell wall can form hole gradually because of decomposition, grease in little algae then can be in the aperture of hole emersion is to the nutrient solution of yeast culture during greater than the grease volume, the stoste that remains in little algae then can be removed after cell walls decomposes fully and the use of classifying.

2. the method for microalgae cell walls broken wall as claimed in claim 1, it is characterized in that, this little algae quantity of adding little algae be can be controlled in allow fungi fully will be little algae still maintain the preset value of fungi quantity after decomposing, so sustainablely after the little algae stoste of taking-up add little algae again to reach a complete circulation process.

3. the method for microalgae cell walls broken wall as claimed in claim 1 is characterized in that, the thalline kind of yeast culture can be used fungi.