	US 12,467,071 B2
	Method for producing secreted β-galactosidase
Eiji Ishikawa, Tokyo (JP); Masakazu Ikeda, Tokyo (JP); Minako Anbe, Tokyo (JP); and Hiroshi Hatano, Tokyo (JP)
Assigned to KABUSHIKI KAISHA YAKULT HONSHA, Tokyo (JP)
Appl. No. 17/293,189
Filed by Kabushiki Kaisha Yakult Honsha, Tokyo (JP)
PCT Filed Nov. 7, 2019, PCT No. PCT/JP2019/043629 § 371(c)(1), (2) Date May 12, 2021, PCT Pub. No. WO2020/100706, PCT Pub. Date May 22, 2020.
Claims priority of application No. 2018-212757 (JP), filed on Nov. 13, 2018.
Prior Publication US 2022/0002771 A1, Jan. 6, 2022
Int. Cl. C12P 19/04 (2006.01); C12R 1/69 (2006.01)

CPC C12P 19/04 (2013.01) [C12R 2001/69 (2021.05)]

3 Claims

1. A method for producing a secreted β-galactosidase, comprising integrating a non-secreted β-galactosidase gene derived from Sporobolomyces singularis and a signal sequence into Aspergillus oryzae to produce a secreted β-galactosidase, and

culturing the Aspergillus oryzae in a CDD medium,

wherein the non-secreted β-galactosidase gene derived from Sporobolomyces singularis and the signal sequence has a sequence of SEQ ID NO: 1, 3 or 5, and

wherein the CDD medium comprises dextrin, glucose, NH₄Cl, KCl, K₂HPO₄, MgSO₄·7H₂O, CuSO₄·5H₂O, FeSO₄·7H₂O, ZnSO₄·7H₂O, MnSO₄·5H₂O, AlCl₃, and MOPS-NaOH buffer pH 7.0.