The invention discloses a 5-aminolevulinic acid producing strain and a construction method thereof. The construction method comprises the following steps of (1) knocking out pxpR, PoxB, ptsG, sucC, sucD and pta genes in
bacillus subtilis to obtain a
bacillus subtilis engineering strain; (2) carrying out double
enzyme digestion treatment on the
plasmid pHT43-J by using Aat II and Sma I, integrating the hemO
gene onto the
plasmid pHT43-J subjected to double
enzyme digestion treatment to obtain a recombinant
plasmid pHT43-J-hemO, then carrying out
enzyme digestion treatment on the recombinant plasmid pHT43-J-hemO by using XbaI and
BamHI, and integrating the rhtA
gene onto the recombinant plasmid pHT43-J-hemO subjected to
enzyme digestion treatment to obtain a first
recombinant expression vector; carrying out double
enzyme digestion on the plasmid pHT01 by using
BamHI and XbaI, and integrating the hemF
gene onto the plasmid pHT01 subjected to double
enzyme digestion treatment, so as to obtain a second
recombinant expression vector; and (3) introducing the obtained first
recombinant expression vector and the obtained second recombinant
expression vector into the
bacillus subtilis engineering bacteria, and constructing to obtain the 5-aminolevulinic
acid producing bacteria. The producing
bacteria provided by the invention improves the yield of 5-aminolevulinic acid.