Constructing a framework carrier to stabilize protein conformation

induce high embedding efficiency

and acquire low mass-transfer resistance is an urgent issue in the development of immobilized enzymes. Hydrogen-bonded organic frameworks (HOFs) have promising application potential for embedding enzymes. In fact

no metal involvement is required

and HOFs exhibit superior biocompatibility

and free access to substrates in mesoporous channels. Herein

a facile

in situ

growth approach was proposed for the self-assembly of alcohol dehydrogenase encapsulated in HOF. The micron-scale bio-catalytic composite was rapidly synthesized under mild conditions (aqueous phase and ambient temperature) with a controllable embedding rate. The high crystallinity and periodic arrangement channels of HOF were preserved at a high enzyme encapsulation efficiency of 59%. This bio-composite improved the tolerance of the enzyme to the acid-base environment and retained 81% of its initial activity after five cycles of batch hydrogenation involving NADH coenzyme. Based on this controllably synthesized bio-catalytic material and a common lipase

we further developed a two-stage cascade microchemical system and achieved the continuous production of chiral hydroxybutyric acid (R-3-HBA).