Method for semiaseptic culturing heterotrophic chlorella

Abstract

The method for making semi-sterile culture of heterotrophic chlorella adopts two steps: first step is sterile culture and second step is open layer-volume culture. Said ivnention can simplify cultureoperation, and can greatly reduce culture cost, and can obtain high-purity heterotrophic chlorella.

Description

technical field

[0001] The invention belongs to algae biological culture technology, in particular to a method for cultivating heterotrophic chlorella under heterotrophic conditions with low cost, high purity and fast growth. Background technique

[0002] Algae have a wide ecological distribution, fast growth and high application value. Chlorella is rich in protein, polysaccharides, lipids, chlorophyll, vitamins, trace elements and some biologically active metabolites, and is widely used in health food, feed, food additives, fine chemicals and raw materials for pharmaceutical preparations. For a long time, Chlorella is not only an excellent experimental material in biological research, but also an object of development and utilization that attracts attention. In addition to the normal growth of Chlorella cells using light energy and carbon dioxide under autotrophic conditions, they can also grow and reproduce using organic carbon sources under heterotrophic conditions, simi...

Examples

Embodiment 1

[0013] Comparing the growth state of heterotrophic algae cells under simple aseptic culture and open culture conditions, the results show (such as figure 1 shown). Chlorella was cultured openly in a non-sterilized 20L glass tank (the culture medium was not sterilized), the growth of heterotrophic cells was very slow, the degree of bacterial contamination was high, and the OD value reflecting the density of heterotrophic cells increased slowly. It ended up only getting to around 3. Chlorella is not easy to be centrifuged and precipitated, which is not conducive to the collection of cell products. If feeding (that is, continuing to supplement nutrients and glucose in the culture solution), it is easier to make bacteria grow in large quantities, and the algae collected by centrifugation are impure and have low biomass. The growth curve of heterotrophic cells in the experimental group after sterilization showed that the OD value reflecting the density of heterotrophic cells incr...

Embodiment 2

[0015] The growth state of chlorella in 10L, 20L bottom bottle (feeding) semi-sterile culture is compared respectively, and the result shows (as figure 2 shown). In the first step, all media and culture vessels are autoclaved. Then, the heterotrophic Chlorella species were subjected to strict aseptic culture in a 400ml ventilated culture bottle at a temperature of 26°C and no requirement on light intensity. When the algae cells grow to the logarithmic phase, after 20-24 hours, transfer the algae cells to a 3L Erlenmeyer flask under aseptic conditions and continue to cultivate them in a light incubator as a 20L mass-cultured algae species. In the second step operation, the algae species in the 3L Erlenmeyer flask were transferred to a 10L or 20L bottom flask under non-sterile conditions (in an open system) for cultivation. The upper and lower openings of the lower bottle are ventilated, the culture temperature changes with the laboratory temperature, and the light intensity ...

technical field

[0001] The invention belongs to algae biological culture technology, in particular to a method for cultivating heterotrophic chlorella under heterotrophic conditions with low cost, high purity and fast growth. Background technique

[0002] Algae have a wide ecological distribution, fast growth and high application value. Chlorella is rich in protein, polysaccharides, lipids, chlorophyll, vitamins, trace elements and some biologically active metabolites, and is widely used in health food, feed, food additives, fine chemicals and raw materials for pharmaceutical preparations. For a long time, Chlorella is not only an excellent experimental material in biological research, but also an object of development and utilization that attracts attention. In addition to the normal growth of Chlorella cells using light energy and carbon dioxide under autotrophic conditions, they can also grow and reproduce using organic carbon sources under heterotrophic conditions, simi...

Examples

Embodiment 1

[0013] Comparing the growth state of heterotrophic algae cells under simple aseptic culture and open culture conditions, the results show (such as figure 1 shown). Chlorella was cultured openly in a non-sterilized 20L glass tank (the culture medium was not sterilized), the growth of heterotrophic cells was very slow, the degree of bacterial contamination was high, and the OD value reflecting the density of heterotrophic cells increased slowly. It ended up only getting to around 3. Chlorella is not easy to be centrifuged and precipitated, which is not conducive to the collection of cell products. If feeding (that is, continuing to supplement nutrients and glucose in the culture solution), it is easier to make bacteria grow in large quantities, and the algae collected by centrifugation are impure and have low biomass. The growth curve of heterotrophic cells in the experimental group after sterilization showed that the OD value reflecting the density of heterotrophic cells incr...

Embodiment 2

[0015] The growth state of chlorella in 10L, 20L bottom bottle (feeding) semi-sterile culture is compared respectively, and the result shows (as figure 2 shown). In the first step, all media and culture vessels are autoclaved. Then, the heterotrophic Chlorella species were subjected to strict aseptic culture in a 400ml ventilated culture bottle at a temperature of 26°C and no requirement on light intensity. When the algae cells grow to the logarithmic phase, after 20-24 hours, transfer the algae cells to a 3L Erlenmeyer flask under aseptic conditions and continue to cultivate them in a light incubator as a 20L mass-cultured algae species. In the second step operation, the algae species in the 3L Erlenmeyer flask were transferred to a 10L or 20L bottom flask under non-sterile conditions (in an open system) for cultivation. The upper and lower openings of the lower bottle are ventilated, the culture temperature changes with the laboratory temperature, and the light intensity ...

Images