Pyruvate stimulates transamination of leucine into α-ketoisocaproic acid and supports 3-methylbutanal production by Lactococcus lactis.

AIM: To investigate the effect of pyruvate and glucose on leucine transamination and 3-methylbutanal production by Lactococcus lactis, including the comparison with cells possessing glutamate dehydrogenase (GDH) activity. METHODS AND RESULTS: Lactococcus lactis cells were incubated in chemically defined medium (CDM) with the pH controlled at 5.2 to mimic cheese conditions. Pyruvate supplementation stimulated the production of the key flavour compound 3-methylbutanal by 3-4 times after 72 h of incubation. Concurrently, alanine production increased, demonstrating the involvement of pyruvate in transamination reactions. Glucose-metabolizing cells excreted α-ketoisocaproic acid and produced even 3 times more 3-methylbutanal after 24 h than pyruvate-supplemented cells. Conjugal transfer technique was used to transfer the plasmid pGdh442 carrying the gdh gene encoding for GDH to L. lactis. Introducing GDH did not stimulate the excretion of α-ketoisocaproic acid and the production of 3-methylbutanal. CONCLUSIONS: These results demonstrate that Lactococcus uses pyruvate to transaminate leucine into α-ketoisocaproic acid which supports 3-methylbutanal production. Surprisingly, GDH activity did not stimulate leucine transamination and 3-methylbutanal production.

Contribution of the novel sulfur-producing adjunct Lactobacillus nodensis to flavor development in Gouda cheese.

We assessed the efficacy of Lactobacillus nodensis CSK964 as an adjunct culture in Gouda cheese under various industrial conditions. We set up 4 different systems: a direct vat inoculum with and without adjunct using the calf rennet Kalase, and an undefined bulk starter culture with and without adjunct using the microbial rennet Milase (both rennets from CSK Food Enrichment, Ede, the Netherlands). During ripening, we subjected the cheeses to the following analyses: viability of starter and adjunct cells, composition, proteolysis, and flavor development by detection of sulfur compounds and descriptive sensory analysis. In general, the presence of Lb. nodensis increased secondary proteolysis and influenced cheese flavor, particularly in relation to volatile sulfur compounds; hydrogen sulfide and methanethiol were present in higher abundances in cheeses containing Lb. nodensis. The primary starter also influenced the range of volatile sulfur compounds produced. Methanethiol and dimethyl disulfide were more abundant in the nisin-producing direct vat inoculum cheese with adjunct; hydrogen sulfide was more prevalent when bulk starter culture was used with Lb. nodensis. Sensory analysis revealed that the direct vat inoculum cheese with adjunct scored significantly better in terms of smell and taste than the direct vat inoculum cheese without adjunct and lacked the dominant sulfur flavors of the bulk starter cheese with adjunct. Subsequent analysis using lead acetate paper and modified motility broth as indicators of hydrogen sulfide production confirmed that Lb. nodensis produced hydrogen sulfide in broth and in the cheese matrix. This study suggests that the inclusion of Lb. nodensis as an adjunct culture can significantly alter the flavor profile of the final cheese. However, the selection of a suitable primary starter is imperative to ensure a desirable product.