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Downstream Process For The Production of Yeast Extract Using Brewer's Yeast Cells
Downstream Process For The Production of Yeast Extract Using Brewer's Yeast Cells
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fkqolar`qflk= ase, and deaminase [6-8]. This process, when using food
proteins such as hydrolyzed vegetable proteins, is gener-
Yeast has been utilized in several areas of food produc- ally composed of two main parts: enzymatic hydrolysis
tion, including brewing, wine, and baking. Brewer’s yeast (upstream process) and the downstream process [9-11].
is an inexpensive nitrogen source, and is generally recog- Downstream processing includes solid-liquid separation,
nized as safe (GRAS). Brewer’s yeast also has good nu- clarification, Maillard reaction, debittering, formulation
tritional characteristics, and contains plenty of protein, concentration, and finally drying. After the yeast hydroly-
lipids, RNA, vitamins, and minerals. sis process, solid particles (yeast cells, macro colloids,
Yeast extract, produced from yeast cells, consists pri- and suspended matter) and haze-forming solutes are re-
marily of amino acids, peptides, nucleotides, and other moved. This is called clarification. Several processes, in-
soluble yeast cell components. Yeast extract can be used cluding membrane filtration and the addition of floccu-
as a flavoring agent in soups, sauces, gravies, stews, lants, can be used to clarify yeast hydrolysate. Addition-
snack foods, and canned foods. Other applications in- ally, a debittering process is required in commercial pro-
clude vitamin supplements in health foods, and nutrients duction, in order to eliminate the bitter and objectionable
in microbiological media [1]. Yeast extract is manufac- taste of yeast hydrolysate. Several debittering processes
tured via cellular breakdown induced by either endoge- using polystyrene divinylbenzene adsorbents have been
nous or exogenous enzymes. Chemical or enzymatic hy- applied in the citrus industry [12]. A combined treatment,
drolysis is the most efficient method for solubilizing yeast. consisting of membrane filtration and the debittering
Enzymatic hydrolysis is accomplished by either prote- process, was investigated in the processing of grapefruit
olytic enzymes [2,3], cell wall lysis enzymes [4] or cul- juice [13]. The debittering of red grapefruit juice was
ture broth containing Streptomyces sp. [5], and produces successfully performed using a dual technique, consisting
yeast extract with a low salt content. of ultrafiltration coupled with an XAD-16 adsorption
We previously reported the enzymatic hydrolysis proc- column [14].
ess for the production of yeast extract, using several In this study, we also report the utilization of waste
kinds of enzymes, including glucanase, protease, nucle- yeast cells from the beer industry in the production of
yeast extract, which can be used as a flavoring foodstuff,
G`çêêÉëéçåÇáåÖ=~ìíÜçê mainly focusing on downstream processing. The effects
qÉäW=HUOJQNJRQMJRSQO= = c~ñW=HUOJOJSOUMJSPQS= of downstream processing on the final product quality of
ÉJã~áäW=ÜàÅÜ~É]çÑÑáÅÉKÜçëÉçK~ÅKâê= the yeast extract were evaluated, including the flavor en-
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hancing step, Maillard reaction, and debittering step, and tested for their efficacy as clarifying agents. The agents
a scheme was proposed to cover the entire process. were added at concentrations of 0.2% (w/w) and 1%
(w/w) in order to accelerate the flocculation of the yeast
hydrolysate (100 g), causing it to settle down. The mix-
j^qbof^ip=^ka=jbqelap= ture of the hydrolysate and agents was stirred at 100 rpm
= for 30 min, then allowed to settle down without mixing
j~íÉêá~äë= = for 30 more min. After the treatment, the mixture (40 g)
was centrifuged at 3,000 × g for 30 min. In order to
Dried brewer’s yeast cells, Saccharomyces sp., were ob- measure the degree of browning (DB), the optical density
tained from a beer factory owned by the Doosan Co. (OD) of the supernatant was measured at 490 nm
(Icheon, Korea). Two types of protease, both manufac- (OD490). The remaining mixture (about 60 g) was then
tured by Novozyme (Bagsvaerd, Denmark), were used in filtered using filter paper (Whatman No. 2), and Celite
protein hydrolysis: Protamex™ (endoprotease) and Fla- 545 (5 g) as a filter-aid. In order to measure the degree
vourzyme™ (exoprotease). These proteases were utilized of clarification (DC), the OD of the filtrate at 600 nm
according to the method described by Chae et al. [6]. (OD600) was monitored. DB and DC were determined
Enzyme RP-1 and deamizyme were obtained from according to the following equations:
Amano Pharmaceuticals (Nagoya, Japan). The synthetic
adsorption resins, Diaion™ HP20 (polystyrene divinyl- OD490 treatment - OD490 control
benzene resin) and Amberlite™ XAD7 (aliphatic acrylic DB = × 100 (%) (1)
polymer resin), were obtained from Mitsubishi Chemical OD490 control
(Tokyo, Japan) and Rohm and Haas (Philadelphia, PA,
treatment
USA), respectively. OD600
DC = (1- control
) ×100(%) (2)
OD600
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natively, clarification using flocculating agents has several
Free amino acid composition was determined by re- advantages, including the simplicity of the process equip-
verse phase chromatography [15] using HPLC (Waters, ment, and low operating costs.
Milford, MA). OD490 and OD600 were measured using a
spectrophotometer (UV mini 1240, Shimadzu, Japan). j~áää~êÇ=oÉ~Åíáçå=
All the experimental data represent mean (average) values
from triplicate measurements. The Maillard reaction has been used to produce foods
that look and taste attractive for thousands of years. The
food industry relies on the application of the Maillard
obpriqp=^ka=afp`rppflk= = reaction to produce many foods, including coffee and
= bakery products [21]. The Hodge scheme remains in
`ä~êáÑáÅ~íáçå= = wide use today [24]. In essence, it is predicated on the
fact that a reducing sugar (like glucose) can condense
Beer is one of the most common products requiring with a compound possessing a free amino group (such as
clarification during its manufacture [16]. Rough beer an amino acid), yielding a condensation product. Various
must be filtered to remove the yeast and colloidal parti- factors influence the Maillard reaction [25], and can be
cles which are responsible for haze. In general, clarifica- divided into food processing and storage variables. These
tion ensures the biological stability of the final products include the nature of the reactants (the composition of
[17]. In order, then, to select the proper flocculating the raw materials), the temperature-time combination
agent and treatment dosage, we evaluated various floccu- used during heating and storage, pH, the water activity of
lating agents by measuring their optical densities at 490 the food, the presence of oxygen or metals, and the pres-
and 600 nm (OD490 and OD600 indicate the degree of ence of any reaction inhibitors, such as sulfur dioxide
browning and clarification, respectively). Among the ma- [26].
terials we tested, aluminum chloride exhibited the highest The extent of the Maillard reaction of the yeast hydro-
effectiveness in lowering OD600 (increasing DC), as is lysate was determined by measuring the OD490, which
shown in Fig. 1. However, aluminum chloride is not gen- indicates the degree of browning (DB). When the yeast
erally accepted to be safe for use as a food additive in hydrolysate was treated at 100oC for 2 h, the OD490 did
Korea. Calcium chloride, however, also exhibited excel- not exhibit a great deal of variation according to the type
lent efficiency with regard to the clarification of the yeast of sugar added, sugar dosage, or glycine addition (data
hydrolysate. Therefore, calcium chloride was selected for not shown). However, the DB exhibited significant varia-
the process, and the treatment dosage was determined to tion according to treatment time at 121oC as shown in
be 1% (w/v) of the hydrolysate mixture, making the DC Fig. 2. When the yeast hydrolysate was treated at 121oC,
about 52%. Membrane filtration is currently considered a significant effect of glucose (Glc) addition on the DB
to be a very promising technology for this purpose, due was observed. Glycine (Gly) addition, however, still ex-
to its ability to perform wine clarification/filtration/hy- erted no significant effects on DB. It has been established
gienization in one single step, in continuous operation that the rate of brown pigment formation is proportional
with clean-in-place (CIP) strategies. Microfiltration (MF) to the square of amino content, reducing sugar concen-
is typically used as a membrane filtraton-based clarifica- tration, and treatment time. The fact that glycine addition
tion process [18-20]. However, this operation is associ- had no effect on this process can be explained by that the
ated presently with prohibitively high costs [21,22]. Alter- fact that the yeast hydrolysate already had plenty of
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Fig. 5. The entire process diagram for yeast extract production. `lk`irpflk= =
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