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We have chosen three his-tagged enzymes for validating this method, d-PhgAT, HeT and Bs GDH

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M13KO7 helper phages (10 11 PFU/mL) were added to each well and incubated for 45 min at 37 C while shaking

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No, relying on mushrooms for B12 is not recommended as they primarily contain inactive corrinoid analogues rather than bioavailable cobalamin