University of Southern Denmark

The quest for umami

Can sous vide contribute?
Clausen, Mathias P.; Christensen, Morten ; Hveisel Djurhuus, Trine; Duelund, Lars;
Mouritsen, Ole G.

Published in:
International Journal of Gastronomy and Food Science

DOI:
10.1016/j.ijgfs.2018.03.002

Publication date:
2018

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Accepted manuscript

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Citation for pulished version (APA):

Clausen, M. P., Christensen, M., Hveisel Djurhuus, T., Duelund, L., & Mouritsen, O. G. (2018). The quest for
umami: Can sous vide contribute? International Journal of Gastronomy and Food Science, 13, 129-133.
https://doi.org/10.1016/j.ijgfs.2018.03.002

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Author’s Accepted Manuscript

The quest for umami: can sous vide contribute?

Mathias P. Clausen, Morten Christensen, Trine

Hveisel Djurhuus, Lars Duelund, Ole G. Mouritsen

www.elsevier.com

PII: S1878-450X(17)30064-1
DOI: https://doi.org/10.1016/j.ijgfs.2018.03.002
Reference: IJGFS93
To appear in: International Journal of Gastronomy and Food Science
Received date: 30 June 2017
Accepted date: 1 March 2018
Cite this article as: Mathias P. Clausen, Morten Christensen, Trine Hveisel
Djurhuus, Lars Duelund and Ole G. Mouritsen, The quest for umami: can sous
vide contribute?, International Journal of Gastronomy and Food Science,
https://doi.org/10.1016/j.ijgfs.2018.03.002
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Scientific Paper

The quest for umami: can sous vide contribute?

Mathias P. Clausen1, Morten Christensen1, Trine Hveisel Djurhuus1, Lars Duelund1, and
Ole G. Mouritsen2

Abstract
Umami is the fifth basic taste that humans during evolution have been primed to seek in
their diet because it signals protein-rich food and easily accessible amino acids. Umami
is elicited by free glutamate and the sensation is enhanced in a synergetic fashion by
free nucleotides, such as inosinate. The content of free glutamate in foodstuff can be
increased by some cooking, ageing, fermentation, and conservation techniques, of
which fermentation is the most powerful. Tenderization by sous vide has during the last
decade become widely popular both in restaurants as well as the home kitchen. The
question arises whether sous vide treatment of meat increases the umami potential by
producing more glutamate. In a pilot study of sous vide preparation of beef tenderloin
we have found somewhat surprisingly that this is not the case. Furthermore, an analysis
of the texture of the meat showed that sous vide does not tenderize the tenderloin meat,
but in fact make it slightly tougher at short preparation times.

Keywords: Umami; Glutamate; Meat; Sous vide; Texture

________________________________________
*Corresponding author: ole.mouritsen@food.ku.dk
1
Department of Chemical Engineering, Biotechnology and Environmental Technology,
University of Southern Denmark, 55 Campusvej, DK-5230 Odense M, Denmark
2
Nordic Food Lab and Section for Design and Consumer Behaviour, Department of
Food Science, University of Copenhagen, 26 Rolighedsvej, DK-1958 Frederiksberg C,
Denmark

Introduction
Our preferences for the various flavours of food, it be taste, aroma, or mouthfeel, are
determined by a complex combination of physiology, culture, tradition, experiences,
and adaptation (Prescott, 2012; Shepherd, 2011; Mouritsen and Styrbæk, 2017).
However, humans’ preference for foodstuff with sweet and umami taste is likely to be a
universal trait (Mouritsen and Styrbæk, 2014) controlled by evolutionary driving forces
that favour species with easy access to calories and protein-rich nutrition. Our
preference for umami (Mouritsen, 2016) is likely to go back to the invention of cooking
along with the hominins’ use of fire to heat-treat food almost 1.9 million years ago
(Wrangham, 2009; Comody et al., 2011). Humans evolved senses that could steer us
towards food that is rich in calories and proteins. The primary tastes, sweet and umami,
do exactly that, respectively (Mouritsen and Styrbæk, 2014).

However, being large molecules carbohydrates and proteins have no taste of their own
since they cannot be detected by the taste receptors, with the possible exception of short
oligosaccharides (Lapis et al., 2016) and small peptides (Maruyama et al., 2012; Kuroda
et al., 2012). Only when broken down into smaller constituents, in particular free amino
acids and sugars, respectively, can they stimulate appropriate taste receptors, such as the
umami receptor. The same holds true for nucleic acids that only have taste when they
are broken down into free nucleotides. Although nucleic acids are not crucial for human
nutrition, the breakdown products of nucleic acids, specifically free 5’-ribonucleotides
like inosinate, guanylate, and adenylate, can elicit umami taste by a particular synergetic
mechanism involving free glutamate as briefly described below.

Cooking and in particular fermenting vegetables and meat, e.g. by endogenous and
exogenous enzymes, can release sugars, free glutamate, and free nucleotides and hence
lead to food and meals enriched in sweetness and umami. In this light, umami could be
seen as a special bonus from the evolution of cooking and the culinary arts, which made
the food not only more nutritious, but also more tasteful (Yamaguchi and Ninomiya,
2000; Mouritsen and Styrbæk, 2014).

Free glutamate (and to a minor extent free aspartate) and free nucleotides enter a
powerful synergic relation due to an allosteric action on the T1R1/T1R3 umami receptor
(Zhang et al., 2008; Mouritsen and Khandelia, 2012). Being highly non-linear, this
synergy implies that small amounts of one component, e.g., inosinate, can enhance the
sensory perception of glutamate manifold (Yamaguchi and Ninomiya, 2000). Only few
types of foodstuffs, such as sun ripe tomatoes and nori (from the red macroalga species

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Pyropia spp.), can by themselves provide both components for umami synergy.
Therefore, usually at least two different ingredients are required (Mouritsen and Styrbæk,
2014). Alternatively, special attention has to be paid as how to prepare the food, e.g., by
heating or fermentation, so that a window opens up for simultaneous presence or
production of free glutamate and free nucleotides. A particular delicate issue is the
possible opposite effects of heat leading to release of taste compounds, but high
temperature may denature the enzymes that could facilitate the breakdown of proteins
and nucleic acids (Ishiwatari et al., 2013).

The original proposal of umami as a fifth basic taste (Ikeda, 2002) was based on the
discovery of substantial amounts of free glutamate in the brown seaweed konbu used for
production of the Japanese soup stock dashi (Ninomiya, 1998; Yoshida, 1998). The
synergetic action in Japanese dashi is mediated by inosinate from a fish product,
katsuobushi (skipjack tuna), and in vegetarian dashi by guanylate from dried shiitake.
Preparation of katsuobushi involves five different techniques: cooking, salting, drying,
smoking, and fermentation by a mold of Aspergillus glaucus. The umami-pairing
principle behind the delicious flavour of dashi is the same that is operative in traditional
western soup stocks prepared by cooking vegetables and meat together.

Preparing fermented fish products, such as fish sauces (Mouritsen et al., 2017), could in
principle provide both glutamate and inosinate, because fresh fish like mackerel,
anchovies, and sardines contain large amount of ATP that enzymatically can be turned
into inosinate, and the subsequent fermentation may produce plenty of glutamate.
However, inosinate is quickly broken down enzymatically soon after the fish is dead
and vanish almost completely during fermentation of fish sauces leading to less-well
tasting compounds, such as inosine or hypoxanthine (Gill, 1990) which is a degradation
product of inosinic acid and have bitter taste (Tikk et al., 2006). The amount of free
inosinate and its degradation derivatives vary during the post-mortem period (Aliani et
al., 2013) and depends on the breed (Koutsidis et al., 2007).

Very special preparation techniques, e.g., as perfected in the Japanese production of

katsuobushi and niboshi (sardines), have to be used in order to conserve the inosinate.
Alternatively the fish could be cooked, dried, and marinated immediately after capture
(Maga, 1983), e.g., for producing anchovy paste. In both cases, however, the proteolytic

3
enzymes that could produce glutamate during subsequent fermentation would be
inactivated, leading to a product with only little free glutamate. Hence it appears that
fermented fish sauces, as well as sauces based on molluscs, insects, pulses, and meat
would predominantly contain free glutamate and very little free nucleotides (Mouritsen
et al., 2017).

When it comes to sous vide cooking of meat it is the same types of considerations that
come into question when optimizing umami flavour and finding a window where
sufficient amounts of free glutamate and free inosinate are present to allow for
synergetic action.

Flavour and sous vide cooking of meat

The recent advances in sous vide techniques (Schellekens, 1996; Myhrvold, 2010;
Baldwin, 2012) have in many respects revolutionized our way of performing accurate
and reproducible culinary preparations of foodstuff, in particular meat (Roldán et al.,
2013). By accurately controlling temperature and cooking time in addition to keeping
the moist as well as soluble taste compounds and the volatile aroma substances of the
foodstuff in or near the cooked item, maximum control over taste, aroma, and texture
can be exercised. In the present paper we shall not be concerned with aroma
compounds, a topic that has received a lot of attention in the context of meat stocks
whose aroma arises as a delicate competition between cooking times and temperature
due to the varied volatility and water solubility of the different aroma compounds
(Snitkjær et al., 2010).

Although flavour is a key issue for sous vide cooking of meat, there appears to be rather
few studies concerned with umami, most of which are considering pork meat (Sasaki et
al., 2007; Ishiwatari et al., 2013; Rotola-Pukkil et al., 2015; Choea et al., 2016; Aaslyng
and Meinert, 2017) and a few beef meat (Mortensen et al., 2015. Cooking temperature
and pH turn out to be more important that cooking time for the overall flavour, in
particular umami in pork (Courts, 1954; Ishiwatari et al., 2013).

In the present paper we present some preliminary results from a pilot study of sous vide
preparation of beef tenderloin focussing on the content of free amino acids, in particular
glutamate. We also present some results for tenderness measured by texture analysis.

4
Although tenderloin is usually not prepared by sous vide cooking since it is already
tender, we choose to study this particular muscle because the fibre structure is rather
homogeneous and with fairly evenly distributed collagen and fatty tissue. This renders
the different samples more similar, although we did observe some variation in the way
the different samples yielded under the texturizer probe.

Materials and methods

Materials
Whole beef tenderloin (psoas major muscle) meat, ca. 2,500 g, was purchased from a
local supermarket (Bilka, A/S, Odense). After slaughtering, the meat has spent two days
in the slaughterhouse and two days in the retail store at 5°C. Before subject to any
treatment in the laboratory, the meat was trimmed and cut crosswise into pieces about
1.5 cm in thickness. The pieces were vacuum packed in heat-stable, food-grade plastic
pouches with each three pieces, representing the thickest, the middle, and the thinnest
part of the tenderloin. The meat was then frozen at -19°C and stored until use. Before
use, the meat was thawed in a refrigerator at 4°C over night.

The acetonitrile for amino-acid analysis was obtained from Fischer scientific
(Walthamn, MA). All other solvent and salts used for chemical analysis were from
Sigma-Aldrich (St. Louis, MO). The water used for chemical analysis was ultra-pure
water obtained from a MilliQ-Integral 5 water purification unit (Merck-Millipore,
Billerica, MA).

Sous vide techniques

The preparation method follows the procedures in earlier work on pork tenderloin
(Choea et al. 2016). We used a La Minerva Pack 10x (Bologna, Italy) and a Julabo TW8
water bath (Seelbach/Germany). Most experiments were performed at 54°C and a
smaller number of experiments were carried out at a higher temperature of 64°C.
Multiple samples were prepared simultaneously and removed from the bath at times t =
0.5, 1, 1,5, 2, 3, 4, 6, and 8 hr. After removal from the water bath, the samples were,
along with an uncooked sample (t=0), subject to texture analysis, and subsequently
stored at -19°C, until used for amino-acid analyses.

Texture analysis

5
After sous vide heat treatment, the sous vide pouches were opened and each piece of
meat was stamped out in a circular form with a diameter of 2.5 cm with a cork borer
(Carl Friedrich Usbeck KG, Radevormwald, Germany). The meat pieces were lightly
dabbed with paper tissue to remove any surface moist and placed in a TA.XT plus
texture analyser (Stable Microsytems, Godalming, United Kingdom) equipped with a 5
mm cylindrical probe (model P/5 from Stable Microsysytems) and a 5 kg load cell
(Stable Microsysytems). The probe was initially placed 20 mm above the middle of the
meat piece. Both the thickest, the middle, and the thinnest part of the tenderloin were
subject to the texture analysis. During the experiment the probe was moved at a speed of
10 mm/s at indentation (and 10 mm/s upon retraction) and was set to move 17 mm into
the meat before retraction. Some other settings were invoked for probing plasticity and
elasticity, where repeated indentation and retraction cycles were implemented. The data
reported below correspond to averages over three measurements made on each of three
samples, i.e., nine measurements.

Amino acid analysis

The methods for extracting amino acids followed earlier published procedures
(Koutsidis et al. 2008; Dermiki et al. 2013). The free amino acids were extracted by
finely blending the meat and then mixing 3 g of this blend with 10 mL 0.1M HCl or
pure water. The mixtures were then shaken for 1 hour at room temperature in a Biosan
E20 (Biosan Riga, Latvia) and then stored at 4ºC over night. This was followed by a
centrifugation for 5 min at 5,000 rpm in an Thermo Scientific CR3i centrifuge (Thermo
Scientific, Waltham, MA, USA) in order to sediment the solid parts. The extracts were
then diluted 20 times with 0.1 mM HCl and subsequently filtered through a 0.2 µm RC
syringe filter (Satorius AG Göttingen, Germany).

Detection and quantification of free amino acids were performed by HPLC-MS using a
Shimadzu LCMS 2020 MS equipped with an electrospray interface (ESI). The HPLC
consisted of a DGU20-A5 in-line degasser, two LC-20AD pumps, a high-pressure
mixer, an SIL20A-HT autosampler, a CTO 10 Column oven, and an SPD-20A UV
detector, all from Shimadzu (Shimadzu, Kyoto, Japan). Separation of the 17 different
free amino acids was performed on an Imtak Intrada amino-acid column (Imtakt
Corporation, Kyoto, Japan) by a gradient of acetonitrile with 0.1% formic acid and 100
mM NH4HCO2, in 21 min. The gradient consisted of first 3 min with 14% NH4HCO2

6
followed by an increase of the NH4HCO2 to 100% during the next 7 min. This condition
was maintained for 2 min where after it was returned to 14% in 2 min and then allowed
to re-equilibrate for the rest of the time. Individual AAs were detected by selected ion
monitoring (SIM) at appropriate m/Z values and were quantified by comparison with a
standard curve prepared from commercial available amino-acid standards from Sigma-
Aldrich (St. Louis, MO).

Results
Texture and tenderness
Representative data from the texture analysis applied to meat cooked at 54°C are shown
in Fig. 1 in case of the middle part of the tenderloin. Each curve corresponds typically to
measurements on nine independent samples. The data for the thin and thick part look
similar (data not shown) with a tendency for the thin end of the tenderloin being to be
more tender than the middle part and the thick end. The trend in the data in Fig. 1
demonstrates that the meat gets progressively less tender the longer the cooking time.
The same trend is found for the samples cooked at 64°C (data not shown).

Since the force curves in Fig. 1 do not exhibit any extended region of linear response we
can conclude that the samples do not behave ideally elastically and a Youngs modulus
cannot easily be determined. In order to get an integral measure of the tenderness we
have therefore integrated the curves in Fig. 1 and similar curves for other conditions to
yield the average work performed on the meat as shown in Fig. 2. It is found that the
work shows an increasing tendency as the cooking time is extended. The average work
is also found to be slightly larger at the higher cooking temperature of 64°C.

In order to more precisely probe the nature of the mechanical response of the sous vide
cooked meat we have subjected the meat pieces to cycles of indentation and retraction.
Representative results are shown in Fig. 3. It is seen that there is only an approximate
linear response at 10 mm indentations, and the lack of elasticity is even more prevalent
at 16 mm indentations, revealing a plastic behaviour of the meat.

Amino acid composition

Some selected and representative results for the glutamate (Glu) contents in samples
extracted with both water or HCl are shown in Fig. 4. It is noted that the content of free

7
Glu appears not to change upon sous vide cooking and that this result is independent on
whether water or HCl extraction methods are used. Approximately the same amount of
Glu is extracted by both techniques. Results for the other free amino acids (data not
shown) demonstrate a similar independence of sous vide cooking and method of
extracting the amino acids. The results of the amino acid composition is an average of
the different pieces of each sample.

Discussion and conclusion

In the present short communication we have first briefly reviewed the question as how
to optimize umami taste in foodstuff, e.g., meat, by various preparation techniques
including fermentation and cooking. We have then raised the question whether the
popular application of sous vide techniques to cooking meat can contribute to those
components of the total flavour experience that pertain to the umami taste and the
tenderness, respectively.

Obviously, culinary experience and a number of scientific studies (Baldwin, 2012,

Myhrvold, 2010) have shown that for various meat cuts, in particular the less tender
meat and meat from older animals, sous vide preparation can in many cases provide for
superior tender, juicy, and very flavourful meat preparations, often with an attribute of
umami. Although, the deliciousness of these preparations is likely to be indisputable it
is unclear whether the actual chemical compounds that elicit umami taste is present and
positively contribute to the deliciousness.

In order to shed some light on this question we have in this paper described the results
of a pilot study of the texture and the contents of umami compounds, i.e., free
glutamate, in sous vide preparations of beef tenderloin. Somewhat surprisingly we
found that the contents of free glutamate is not changed by sous vide cooking at
temperatures 54°C or 64C° and hence the umami taste is the same as that of raw meet. It
should be pointed out that our analysis is based on measuring free amino acids in the
total sample and not only of the juices released during cooking, in which it may well be
that free glutamate is increased over time as found in studies of sous vide cooked pork
(Rotola-Pukkila et al., 2015).

8
Regarding texture and tenderness we find that the meat gets slightly less tender with
increased cooking time and increased temperature. The mechanical response of the meat
is complex and there is no elastic regime discernable. There is some indication that the
thin end of the tenderloin is slightly tenderer than the middle and thick part. These
results are most likely peculiar to our choice of tenderloin, and sous vide preparation of
more tough muscles would show very different dependence of time and temperature
(Tornberg, 2005).

As shown in Fig. 4, the typical free glutamate content in the sous vide cooked beef
tenderloin samples are around 10 mg/100 g which is somewhat lower that some
literature values for untreated beef meat, about 30 mg/100 g (Maga, 1983; Ishiwatari et
al., 2013;) as well as for sous vide data for semitendinosus of beef muscle (Jorge Ruiz
Carrascal, personal communication). These variations are most likely to be caused by
the differences in the chosen meat cut and the extraction methods used.

The results presented in this paper do not imply that the sous vide cooking does not
change the flavour of the meat, and that this preparation techniques in fact can render
the meat more delicious and juicy. Similarly, the results do also not imply that sous vide
techniques are not useful for preparing more tender and juice meat for other types of
meat with more collagen and fat content. Only a full sensory investigation of a range of
meat cuts can uncover this question (Mortensen et al., 2012; 2015). It should be
emphasized that we have only investigated taste as far as it may be elicited by free
amino acids. It is possible, and very likely, that other taste compounds as well as aroma
compounds are developed during sous vide cooking (Baldwin, 2012) rendering the
overall result more delicious and flavourful. Only a full chemical analysis, also
including measurements of free nucleotides, combined with a sensory investigation can
uncover these questions.

Acknowledgments
This work was supported in part by a grant to Smag for Livet (Taste for Life) from
Nordea-fonden (both to University of Southern Denmark and Nordic Food Lab at
University of Copenhagen). The HPLC-MS equipment was supported by a grant from
the VILLUM Foundation. The TA.XTplus texture analyser was supported by a grant

9
from the Carlsberg Foundation. ). We wish to thank Jorge Ruiz Carrascal for useful
information regarding sous-vide studies of beef meat.

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Figures

Fig.1 Force versus time (distance) at indenting probe speed of 1 mm/s for the middle
part of beef tenderloin prepared at 54°C by sous vide cooking for a range of time
periods.

13
Fig. 2 Average integrated work of force-time (distance) curves like in Fig. 1 for probe
speed of 1 mm/s for the middle part of beef tenderloin prepared at 54°C by sous vide
cooking for a range of time periods. The data bar to the right is for a sample cooked
sous vide for 0.5 hr at 54°C.

Fig. 3 Force versus time (distance) at indenting probe speed of 1 mm/s and retraction
speed 10 mm/s for the middle part of beef tenderloin prepared at 54°C by sous vide
cooking for 0.5 hr. Results are shown for total indentations of 10 mm (a) and 16 mm
(b), respectively. The figure shows a succession of ten indentations on the same sample.

Fig. 4 Concentration of free glutamate in beef tenderloin prepared at 54°C by sous vide
cooking for a range of time periods. Results are shown for both water (a) and HCl (b)
extraction.

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