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4 protocols using yeast powder

1

Synthesis of GABA-producing Lactobacillus

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Polyvinyl alcohol (DP: 1750 ± 500), sodium alginate, boric acid, calcium chloride (CaCl2), sodium acetate, acetic acid, and tetrahydrofuran were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). While the tryptone, yeast powder, and L-sodium glutamate (L-MSG) were purchased from Sangon Biotech Co., Ltd. (Shanghai, China) and the chromatographic grade methanol was purchased from Oceanpak Alexative Chemical., Ltd. (Goteborg, Sweden). GABA producing Lactobacillus brevis CGMCC 1306 was isolated in our laboratory from fresh milk.
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2

Synthesis of GABA-producing Lactobacillus

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Polyvinyl alcohol (DP: 1750 ± 500), sodium alginate, boric acid, calcium chloride (CaCl2), sodium acetate, acetic acid, and tetrahydrofuran were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). While the tryptone, yeast powder, and L-sodium glutamate (L-MSG) were purchased from Sangon Biotech Co., Ltd. (Shanghai, China) and the chromatographic grade methanol was purchased from Oceanpak Alexative Chemical., Ltd. (Goteborg, Sweden). GABA producing Lactobacillus brevis CGMCC 1306 was isolated in our laboratory from fresh milk.
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3

Yeast Genomic DNA Extraction Protocol

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Restriction enzymes, T4 DNA ligase, and DNA polymerase were purchased from Takara Biomedical Technology Co., Ltd. (Beijing, China). EasyPure Plasmid Miniprep Kit was purchased from Tsingke Biotechnology Co., Ltd. (Beijing, China). TIANquick Midi Purification Kit was purchased from Tiangen Biotech Co., Ltd. (Beijing, China). Rapid Yeast Genomic DNA Isolation Kit, yeast powder, peptone, kanamycin, ampicillin, G418 sulfate, and YNB (yeast nitrogen base, without amino acid and ammonium sulfate) were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). GlcNAc was purchased from Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). All chemicals used were of analytical grade.
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4

Controlled Error-prone PCR for Mutagenesis

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Restriction enzymes, plasmid extraction kit, and fragment purification kit were purchased from TAKARA (Dalian, China); Controlled Error-prone PCR Kit was purchased from TIANDZ (Beijing, China); ClonExpress II One Step Cloning Kit was purchased from Vazyme (Nanjing, China); yeast powder, peptone, Kanamycin, Ampicillin, Isopropyl-β-d-thiogalactoside, and l-aspartic acid were purchased from Sangon (Shanghai, China); 1,2-Diacetylbenzene was from Sigma-Aldrich (St. Louis, MO, USA). All of the chemicals used were of analytical grade.
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