Overexpression of Lactobacillus casei D-hydroxyisocaproic acid dehydrogenase in cheddar cheese.
Metabolism of aromatic amino acids by lactic acid bacteria is an important source of off-flavor compounds in Cheddar cheese. Previous work has shown that alpha-keto acids produced from Trp, Tyr, and Phe by aminotransferase enzymes are chemically labile and may degrade spontaneously into a variety of off-flavor compounds. However, dairy lactobacilli can convert unstable alpha-keto acids to more-stable alpha-hydroxy acids via the action of alpha-keto acid dehydrogenases such as d-hydroxyisocaproic acid dehydrogenase. To further characterize the role of this enzyme in cheese flavor, the Lactobacillus casei d-hydroxyisocaproic acid dehydrogenase gene was cloned into the high-copy-number vector pTRKH2 and transformed into L. casei ATCC 334. Enzyme assays confirmed that alpha-keto acid dehydrogenase activity was significantly higher in pTRKH2:dhic transformants than in wild-type cells. Reduced-fat Cheddar cheeses were made with Lactococcus lactis starter only, starter plus L. casei ATCC 334, and starter plus L. casei ATCC 334 transformed with pTRKH2:dhic. After 3 months of aging, the cheese chemistry and flavor attributes were evaluated instrumentally by gas chromatography-mass spectrometry and by descriptive sensory analysis. The culture system used significantly affected the concentrations of various ketones, aldehydes, alcohols, and esters and one sulfur compound in cheese. Results further indicated that enhanced expression of d-hydroxyisocaproic acid dehydrogenase suppressed spontaneous degradation of alpha-keto acids, but sensory work indicated that this effect retarded cheese flavor development.
Western Dairy Center, Department of Nutrition and Food Sciences, Utah State University, Logan, UT 84322-8700, USA. broadbnt@cc.usu.edu
Jeffery R
JR
Broadbent
Sanjay
S
Gummalla
Joanne E
JE
Hughes
Mark E
ME
Johnson
Scott A
SA
Rankin
Mary Anne
MA
Drake
D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) / Q000235:genetics / Q000378:metabolism*; D002208:Caproates / Q000737:chemistry / Q000378:metabolism*; D002611:Cheese / Q000382:microbiology*; D003470:Culture Media; D008401:Gas Chromatography-Mass Spectrometry; D007780:Lacticaseibacillus casei / Q000201:enzymology* / Q000235:genetics; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics; D010957:Plasmids; D013649:Taste
D002208:Caproates; D003470:Culture Media; C034527:isocaproic acid; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)