The invention discloses an hph resistance
gene knockout box based on gme5
gene editing, a
genetically engineered bacterium and application of the hph resistance
gene knockout box and the
genetically engineered bacterium. The hph resistance
gene knockout box comprises a
recombinant DNA construction body with the
nucleotide sequence shown as SEQ ID NO: 1, the
recombinant DNA construction body replaces an endogenous regulatory factor gme5 gene through
homologous recombination in a targeted mode, and the
nucleotide sequence of the regulatory factor gme5 gene is shown as SEQ ID NO: 2. According to the application, the
bumblebee candida albicans are directionally modified through a
genetic engineering strategy, an
engineering strain for synthesizing the high-yield
lactone type
sophorolipid is constructed, and the
lactonase activity and
fatty acid transport capacity are synergistically enhanced, so that the ratio of the
lactone type
sophorolipid is maximized, and the yield of the
lactone type
sophorolipid is increased. In the sophorolipid produced by using the
genetically engineered bacterium and the
fermentation production method disclosed by the invention, the total content of the sophorolipid is 358.77 g / L, and the lactone content accounts for 74% or more of the total sophorolipid, so that the separation and purification difficulty and the production cost are greatly reduced, and an efficient and low-cost solution is provided for
industrial fermentation production of the sophorolipid.