Ballerup, Denmark
Ballerup, Denmark

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Crafack M.,Copenhagen University | Mikkelsen M.B.,Copenhagen University | Saerens S.,Chr. Hansen A S | Knudsen M.,Copenhagen University | And 7 more authors.
International Journal of Food Microbiology | Year: 2013

The potential impact of aromatic and pectinolytic yeasts on cocoa flavour was investigated using two defined mixed starter cultures encompassing strains of Pichia kluyveri and Kluyveromyces marxianus for inoculating cocoa beans in small scale tray fermentations. Samples for microbial and metabolite analysis were collected at 12-24hour intervals during 120h of fermentation. Yeast isolates were grouped by (GTG)5-based rep-PCR fingerprinting and identified by sequencing of the D1/D2 region of the 26S rRNA gene and the actin gene. Pulsed Field Gel Electrophoresis (PFGE) was conducted on isolates belonging to the species P. kluyveri and K. marxianus to verify strain level identity with the inoculated strains. Furthermore, Denaturing Gradient Gel Electrophoresis (DGGE) was performed to follow yeast and bacterial dynamics over time including the presence of the bacterial inoculum consisting of Lactobacillus fermentum and Acetobacter pasteurianus. Yeast cell counts peaked after 12h of fermentation with the predominant species being identified as Hanseniaspora opuntiae and Hanseniaspora thailandica. P. kluyveri and K. marxianus were found to compose 9.3% and 13.5% of the yeast population, respectively, after 12h of fermentation whilst PFGE showed that ~88% of all P. kluyveri isolates and 100% of all K. marxianus isolates were identical to the inoculated strains. Despite never being the dominant yeast species at any stage of fermentation, the un-conched chocolates produced from the two inoculated fermentations were judged by sensory analysis to differ in flavour profile compared to the spontaneously fermented control. This could indicate that yeasts have a greater impact on the sensory qualities of cocoa than previously assumed. © 2013 Elsevier B.V.

Klindt-Toldam S.,Copenhagen University | Larsen S.K.,Copenhagen University | Saaby L.,Copenhagen University | Olsen L.R.,Toms Confectionary Group A S | And 6 more authors.
LWT - Food Science and Technology | Year: 2016

Probiotic microorganisms are exposed to different types of stress both before and after consumption which strongly influences probiotic survival. Here the viability of 2 different probiotic bacteria incorporated in different types of chocolate was investigated during storage and passage of the upper gastro-intestinal tract using both a static and a dynamic gastric in vitro model. Lactobacillus acidophilus NCFM® and Bifidobacterium lactis HN019 equalling a total concentration of 2 × 108, 2 × 109 and 2 × 1010 CFU/g chocolate were added to samples of milk chocolate, 57% and 72% dark chocolate, respectively. The probiotic strains tolerated the manufacturing process reasonable well with a reduction of 1.1–1.6 log CFU/g after one month of storage, and showed an excellent survivability during 14 months of storage, especially when kept at a constant temperature of 15 °C. In comparison with commercial dairy and juice based probiotic products, chocolate was an excellent carrier for probiotic delivery, because of very good survival of probiotics during simulated passage of the upper GI tract. The viability of B. lactis was slightly higher than Lb. acidophilus and survival rates were >6.5 log CFU/g in case of the static, and >7.0 log CFU/g, in case of the dynamic gastric model, with milk chocolate being the most protective carrier. © 2016 Elsevier Ltd

Crafack M.,Copenhagen University | Keul H.,Copenhagen University | Eskildsen C.E.,Copenhagen University | Petersen M.A.,Copenhagen University | And 7 more authors.
Food Research International | Year: 2014

The sensory quality of chocolate is widely determined by the qualitative and quantitative composition of volatile compounds resulting from microbial metabolism during fermentation, and Maillard reactions taking place during drying, roasting and conching. The influence of applying mixed starter cultures on the formation of flavour precursors, composition of volatile aroma compounds and sensory profile was investigated in cocoa inoculated with cultures encompassing a highly aromatic strain of Pichia kluyveri or a pectinolytic strain of Kluyveromyces marxianus, and compared to commercially fermented heap and tray cocoa. Although only minor differences in the concentration of free amino acids and reducing sugars was measured, identification and quantification by dynamic headspace gas chromatography-mass spectrometry (HS/GC-MS) revealed pronounced differences in the composition of volatiles in roasted cocoa liquors and finished chocolates. 19 of the 56 volatile compounds identified in the chocolates were found in significantly higher amounts in the tray fermented sample, whilst significantly higher amounts of 2-methoxyphenol was measured in the two inoculated chocolates. The P. kluyveri inoculated chocolate was characterized by a significantly higher concentration of phenylacetaldehyde and the K. marxianus inoculated chocolate by significantly higher amounts of benzyl alcohol, phenethyl alcohol, benzyl acetate and phenethyl acetate compared to a spontaneously fermented control. Sensory profiling described the heap and tray fermented chocolates as sweet with cocoa and caramel flavours, whilst the inoculated chocolates were characterized as fruity, acid and bitter with berry, yoghurt and balsamic flavours. The choice of fermentation technique had the greatest overall impact on the volatile aroma and sensory profile, but whilst the application of starter cultures did affect the volatile aroma profile, differences were too small to significantly change consumer perception of the chocolates as compared to a spontaneously fermented control. © 2014 Elsevier Ltd.

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