To explore the potential application of edible seaweed in traditional fermented foods, Gracilaria was used as the research object, and the response surface methodology was used to optimize the koji-making process of soy sauce by adding Gracilaria.The effects of adding Gracilaria powder on the spore count, enzyme activity and solid content in the soy sauce koji making process of were investigated, and further gas chromatography-ion migration spectrometrycombined with principal component analysis were used to analyze the volatile flavor compounds of traditional soy sauce koji and Gracilaria koji.Results showed that the optimum Gracilaria koji making conditions were that koji-making time was 86 h, the ratio of Aspergillus oryzae Aspergillus Niger was 1∶3 with a inoculation amount of 4.3%, the ratio of soybean powder,flour, and Gracilaria was 5∶2∶2.5.Under these conditions, the acid protease activity was 2 956.86 (U/g dry basis) and glucoamylase activity was 3 082.80 (U/g dry basis).The activities of acid protease, glucoamylase, amylase, aminopeptidase, and cellulase and the number of spores in Gracilaria koji were higher than those in traditional soy sauce koji, and the solid content was lower than that in soy sauce koji.A total of 24 flavor substances were identified in traditional soy sauce koji and Gracilaria koji.The addition of Gracilaria affected the types and content of volatile flavor substances in soy sauce koji, among which alcohols, ketones, esters, and aldehydes contributed more to the flavor of Gracilaria koji.GC-IMS combined with principal component analysis could distinguish the main flavor substances of traditional soy sauce koji and Gracilaria koji.In summary, the addition of Gracilaria in the koji-making process can significantly improve its enzyme activity and endow the system with different flavor substances, indicating Gracilaria has the potential to brew soy sauce as a fermentation substrate, and providing theoretical reference for the high-value utilization of seaweed resources and enriching the variety of soy sauce products.
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