The invention discloses a preparation method of a novel composite
carbon source. The preparation method comprises the following steps: S1, preparing a solution; s2, preparing a culture medium; s3, extracting a
genome; s4, extraction of
plasmid DNA (deoxyribonucleic acid); s5, preparation of
agarose gel electrophoresis; s6, constructing a linearized vector
plasmid; s7, preparing an electrotransfection
competent cell; and S8, electric conversion treatment. According to the preparation method of the novel composite
carbon source, pfkA and pfkB can be respectively, independently and simultaneously knocked out when the
carbon source is prepared by utilizing a
gene editing technology, the consumption of a
glucosamine precursor 6-
fructose phosphate in an EMP pathway can be reduced, more glucose is used for synthesizing a target product, and by optimizing the preparation process of the composite carbon source, the yield of the target product is improved. The shake-flask
fermentation yield of the
glucosamine can be obviously improved, the conversion rate of the composite carbon source is also improved, and
cell growth and synthesis of the target product
glucosamine can be reasonably balanced, so that the yield of the glucosamine is improved.