CN106479895A - A kind of method of utilization xylose Combined hardening model chlorella - Google Patents

Abstract

The invention discloses a kind of method of utilization xylose Combined hardening model chlorella, it is characterized in that a kind of bacterium algae co-culture system is constructed with antibacterial and chlorella, within the system, antibacterial carries out growth metabolism with xylose for carbon source, and its metabolite provides organic carbon source for the mixotrophic growth of chlorella；Using the method, chlorella cells growth rate is more than 3 times of its photoautotrophy growth.The present invention is greatly facilitated the growth of chlorella, shortens the time of illumination cultivation, decreases the energy loss during illumination cultivation, can provide big content of starting materials for the industrialized production of microalgae biodiesel simultaneously.

Description

A kind of method of utilization xylose Combined hardening model chlorella

Technical field

The present invention relates to chlorella culture technique field is and in particular to a kind of utilization xylose Combined hardening model chlorella Method, the frustule of results can be that the production of biodiesel provides cheap raw material.

Background technology

Biodiesel is a kind of green reproducible energy, is subject to the common concern of countries in the world in recent ten years.At present, Biodiesel is mainly derived from plant and animal oils and fatss, and countries in the world select suitable raw material, EU countries according to itself national conditions With Oleum Brassicae campestriss as raw material, the U.S., Brazil (the third place in the world mcroorganism diesel oil place of production, 2011 2500000 tons) of South America and Ah The root court of a feudal ruler (the world the fourth-largest biodiesel place of production, 2011 2300000 tons) main with soybean oil as raw material, country in Southeast Asia is main Produce biodiesel using Petiolus Trachycarpi oil, Japan is then with kitchen waste oil as primary raw material.Stable glyceride stock supply is current state One of fast-developing key of outer Biodiesel, but existing yield also only has 5% market part in the first big producing region European Union Volume, the development further of biodiesel will consume substantial amounts of glyceride stock, and this will result in huge to the sound development of agricultural and forestry Big pressure；Especially in the rare China of populous, per capita area of cultivated farmland, Semen sojae atricolor and grain and oil rely primarily on import, biological bavin The development of oil need under the principle of " do not strive with the people grain and oil, do not strive with grain and oil ground " active development non-grain and oil raw material sources.In the face of planting Raw material produces the problems of biodiesel, and microalgae biodiesel is increasingly paid attention to by various countries.Chlorella is a class list Cell algae, belongs to Chlorophyta, and species is various, all has widely distributed in fresh sea water, also can under culture conditions Good growth, is the microalgae of the first artificial culture.Because chlorella distribution is wide, growth is fast, the advantages of easily culture, become production The important source material source of biodiesel.

In lignocellulose ~ 30% be hemicellulose, the 85 ~ 90% of hydrolysis of hemicellulose product is xylose.Xylose is as one Plant fermentable sugars, its fermentation producing and ethanol can make the ethanol production of cellulosic material improve more than 25% in original basis, in cellulose Get most of the attention in ethanol industry.In the culture of microalgae, xylose as carbon source carry out heterotrophism or Combined hardening model research and Application is also seldom.The research of Yang et al. findsScenedesmus obliquuMixotrophism life can be carried out for carbon source with xylose Long, but heterotrophic growth can not be carried out, and when the low xylose concentration of concentration of algae is high, frustule growth is suppressed, to after 10d Start with xylose（Sequential heterotrophy-dilution-photoinduction cultivation for efficient microalgal biomass and lipid production. Bioresource Technology, 2012, 112: 206-211.）.Zheng et al. research it was also found that by induction (microalgae is first cultivated in dextrose culture-medium, The cell obtaining as has received the cell of induction)Chlorella sorokinianaFermenting xylose very capable, and not The cell ability of induction is but very weak；They speculateC. sorokinianaThere is a kind of hexose carrier, this carrier in cell After being induced activation, xylose just can be carried entrance frustule, be converted into xylulose by Xylose reductase and xylitol dehydrogenase and enter Enter phosphopentose pathway（NAD(P)H-linked xylose reductase and NADP+-linked xylitol dehydrogenase in the oleaginous microalgaChlorella sorokiniana. Biotechnology for Biofuels, 2014, 7: 125.）.Comprehensive existing result of study, xylose is to microalgae cell Growth has certain inhibitory action, and frustule has to pass through induction or long adaptation could be mixed for carbon source using xylose Conjunction is nourished and grown.Therefore, research and utilization xylose promotes the method for chlorella growth to saving microalgae production cost, develops biological bavin Oil industry is significant.

Content of the invention

For the feature of chlorella photoautotrophy growth is slow, using the growth characteristics of the facultative chemoheterotrophy of chlorella, this Bright provide a kind of method that utilization xylose is mixed chlorella, a kind of bacterium algae is constructed with antibacterial and chlorella and co-cultures system System, within the system, antibacterial carries out growth metabolism with xylose for carbon source, and its metabolite is that the mixotrophic growth of chlorella carries For organic carbon source.This method is greatly facilitated the growth of chlorella, shortens the time of illumination cultivation, decreases illumination cultivation mistake Energy loss in journey, can provide big content of starting materials for the industrialized production of microalgae biodiesel simultaneously.

The invention provides a kind of method of utilization xylose Combined hardening model chlorella, the method is with antibacterial and chlorella Construct a kind of bacterium algae co-culture system, within the system, antibacterial carries out growth metabolism with xylose for carbon source, metabolite is little The Combined hardening model of ball algae provides organic carbon source.

Further, antibacterial used is Cellvibrio Cellvibrio pealriver, protects in Chinese microorganism strain The preserving number hiding administration committee's common micro-organisms center is CGMCC 1.14955, and preservation date is on December 21st, 2014.

Further, chlorella used is Chlorella vulgaris.

Further, co-culture system refers to that Cellvibrio pealriver and Chlorella vulgaris connects simultaneously Plant in culture medium.

Further, the culture medium that bacterium algae co-cultures is to add xylose in BG11 culture medium for carbon source.

Further, Combined hardening model mode refer to simultaneously with Cellvibrio pealriver xylose metabolism product and CO2 is carbon source illumination cultivation Chlorella vulgaris.

Compared with prior art, the present invention has the advantage that and technique effect：

1, the invention provides a kind of method of utilization xylose mixed culture chlorella, constructs a kind of bacterium with antibacterial and chlorella Algae co-culture system, within the system, antibacterial carries out growth metabolism with xylose for carbon source, and its metabolite is the mixing of chlorella Nourish and grow offer organic carbon source.

2nd, the method for the present invention is greatly facilitated the growth of chlorella, shortens the time of illumination cultivation, decreases illumination Energy loss in incubation, can provide big content of starting materials for the industrialized production of microalgae biodiesel simultaneously.

Brief description

Fig. 1 isChlorella vulgarisWhen BG11, xylose and co-culture system growChlA change is bent Line chart (solid line), andC. pearlriverConcentration of reduced sugar change (dotted line) curve chart when growing in xylose.

Specific embodiment

With reference to embodiment, the technology contents of the present invention are further described；Following embodiments are illustrative, It is not determinate it is impossible to limit protection scope of the present invention with following embodiments.

Embodiment one：

The xylose adding 1g/L in BG11 culture medium co-cultures culture medium for bacterium algae, by 1%（v/v）Inoculum concentration inoculate respectivelyCellvibriopealriverWithChlorella vulgarisFACHB-8（Buy from Wuhan Chinese Academy of Sciences Wildlife Germplasm Bank algae kind storehouse）Co-cultured；Matched group respectively with 1. BG11 and 2. BG11+1g/L xylose as culture medium Culture chlorella.5ml culture fluid is taken to extract algae chlorophyll a in incubation daily（Chla）, measurement result is as shown in figure 1, train Frustule fast-growth after foster 2d, its growth rate is 3 times of its photoautotrophy growth, 5d, microalgae in co-culture systemChl A reaches 7.75mg/L；Contrast matched group result 1. and 2., xylose has certain inhibitory action to chlorella growth；To cultivating In journey, the measurement of content of reducing sugar shows, chlorella can not be individually with xylose, and in co-culture system, content of reducing sugar is anxious Acute decline, to depleted during 4d.Summary result we understand, using bacterium algae co-culture system, xylose may quiltC. pearlriverIt is converted into a kind of promotionC. vulgarisThe material of growth, carries out Combined hardening model with xylose for carbon source The growth of chlorella can be significantly facilitated.

Embodiment two：

The xylose adding 5g/L in BG11 culture medium co-cultures culture medium for bacterium algae, by 1%（v/v）Inoculum concentration inoculate respectivelyCellvibriopealriverWithChlorella vulgarisFACHB-8 is co-cultured；Matched group is respectively with 1. BG11 2. BG11+5g/L xylose is culture medium culturing chlorella.5ml culture fluid is taken to extract algae chlorophyll a in incubation daily （Chla）, measurement result shows, cultivates frustule fast-growth after 2d, and its growth rate is the 3 ~ 4 of its photoautotrophy growth Times, 5d, microalgae in co-culture systemChlA reaches 10.21mg/L, than 2.76 mg/L of the growth of photoautotrophy in BG11 Improve 3.7 times.The concentration that the above results explanation improves xylose is conducive to improving the accumulation of chlorella cells.

Embodiment three：

The xylose adding 1g/L in BG11 culture medium co-cultures culture medium for bacterium algae, by 5%（v/v）Inoculum concentration inoculationCellvibriopealriverInoculate with 1% inoculum concentrationChlorella vulgarisFACHB-8 is co-cultured；Comparison Group respectively with 1. BG11 and 2. BG11+1g/L xylose for culture medium culturing chlorella.5ml culture fluid is taken daily in incubation Extract algae chlorophyll a（Chla）, measurement result shows, cultivates frustule fast-growth after 1d, and its growth rate is that it is photosynthetic certainly 2 ~ 3 times of health preserving length, 5d, microalgae in co-culture systemChlA reaches 7.21mg/L, than the growth of photoautotrophy in BG11 2.81mg/L improves 2.6 times.The above results explanation improvesCellvibriopealriverInoculum concentration be conducive to shorten bead Algae adapts to the time of environment, reduces the time of chlorella culture.

Example IV：

The xylose adding 1g/L in BG11 culture medium co-cultures culture medium for bacterium algae, by 1%（v/v）Inoculum concentration inoculationCellvibriopealriverInoculate with 5% inoculum concentrationChlorella vulgarisFACHB-8 is co-cultured；Comparison Group respectively with 1. BG11 and 2. BG11+1g/L xylose for culture medium culturing chlorella.5ml culture fluid is taken daily in incubation Extract algae chlorophyll a（Chla）, measurement result shows, cultivates frustule fast-growth after 1d, and its growth rate is that it is photosynthetic certainly 2 ~ 3 times of health preserving length, 5d, microalgae in co-culture systemChlA reaches 8.14mg/L, than the growth of photoautotrophy in BG11 2.97mg/L improves 2.7 times.The inoculum concentration that the above results explanation improves chlorella is conducive to shortening frustule adaptation environment Time, reduce the time of chlorella culture.

Claims (6)

1. a kind of method of utilization xylose Combined hardening model chlorella is it is characterised in that the method is with antibacterial and chlorella structure Build a kind of bacterium algae co-culture system, within the system, antibacterial carries out growth metabolism with xylose for carbon source, metabolite is bead The Combined hardening model of algae provides organic carbon source.

2. a kind of method of utilization xylose Combined hardening model chlorella according to claim 1 is it is characterised in that institute Antibacterial is CellvibrioCellvibriopealriver, in China Committee for Culture Collection of Microorganisms's commonly micro- life The preserving number at thing center is CGMCC 1.14955, and preservation date is on December 21st, 2014.

3. a kind of method of utilization xylose Combined hardening model chlorella according to claim 1 is it is characterised in that institute Chlorella isChlorella vulgaris.

4. a kind of method of utilization xylose Combined hardening model chlorella according to claim 1 is it is characterised in that be total to Culture systems refer toCellvibriopealriverWithChlorella vulgarisIt is inoculated in culture medium simultaneously.

5. a kind of method of utilization xylose Combined hardening model chlorella according to claim 1 is it is characterised in that bacterium algae The culture medium co-culturing is to add xylose in BG11 culture medium for carbon source.

6. a kind of method of utilization xylose Combined hardening model chlorella according to claim 1 is it is characterised in that mix Joint venture is supported training method and is referred to simultaneously with Cellvibrio pealriver xylose metabolism product and CO2 for carbon source illumination cultivation Chlorella vulgaris.