Characteristic of chanic n Oi rupt d & k r ' f

Yeastin Relation to it parati n in lndu trial

entrifu

rn
F.G. MOSQUIRA,' J. J. Hl '\ .t P I
LiLLY,Department of Chemin.1 lind 8iodlemiClli -II ,jtlte i l1.
UniversilJ College London, forrinl/,on PIlI(' . LamlclI' W I 7J.
En/llolld

ummaf"}'
1 :v.iscosity,density. and sedimenlallon chan.c,en'IIC' of "pe,ns,on of ....hOle l.,nd
.' h , isrupte yeast cells were measured. Mecham al d"rupllon In re'''' the "-
m d'I y an dItSnon.New,on .. n beh.vior. Expenmenl
beteCllamcally ho"ed good com:lnllon
""nSlon VISCOS"t d. . atlon result .
weenlaborat ory- and ..mdusmal-sca1c centnfug
.'

lNTROD CT10

lioMet~odS are. now available for the indu trial-scale mechanical di rup-
di; 0 microbial cells 1.2 to isolate intracellular product . Following cell
recov norma Y necessary to remove the cell debfls for product
uption it is II .
ery.
ru:~dhave previously examined the clarification of mechanical1y di -
at
I fugal'yeast suspensions by rotary vacuum preco filtration.' Centri-
direc~o~ IS the currently used alternative to filtration and this study is
Pe~ toward a better understanding of the use of this method.
I depen~rmancepredictions of solid-liquid separation by these methOds
andth ~n a knowledge of certain physical characteristicS of the solids
farwh~l~qUldSm which they are suspended. A little information exists
simpl cell suspensIOns.'" We have measured several properties usmg
a sap~and readily available techniques that, although they do not justify
ThislslIcatedanalysis," do provide some useful information.
ands laper first describes laboratory studies on the viscosity, density,
e imentation characteristics of disrupted bakers' yeast suspensions

Mexico ress. Centro de Investigacion deiIPN. APto, postal 14-740, Mexico 14, D.F.,
'* Present add
land, ress: New Zealand Dairy Research Inslllule, Palmers!on North, New Zea-
t Present add .

335_343 (1981)
0006-359218110023-0335$01.00
Biotechnolo .
It) \98\ JOh;Y ~nd Bioengineering, Vol.
Wiley & Sons, Inc.
336
MO Q IRA TAL. CHARACTERtSTtCS OF DtSRUPTED BAKERS' YEAST 337

and, secondly, some re ult wasdetermined at SOCby u ing 10011 pycnometers with a perforated plug.
indu trial- cale centrifuge for the edimentation of debri in pilot- and
Proteinwas measured by the Folin-Lowry method.
Laboratory centrifugal sedimentation was carried out at YC in a high-
speedrefrigerated centrifuge (MSE Ltd., Crawley, West Sussex) with an
MATERIALS AND METHODS 8 x 5 mJ swing-out head (MSE model o. 69084) using polypropylene
tubes.Four ml suspension were placed in the tube, which was then cen-
Yeast was obta',ned f rorn The D' '11
:eparately wrapped I lb packs rsu ers Company Ltd. (London), in tnluged under the appropriate conditions of speed and time. Samples of
alOlOg 250mM sodium chi id The yeast was suspended in a buffer con- 1mlwere taken from the centrifuge tube at a fixed depth below the surface
phate with a stirrer-homo 0:' .e and 4mM dipotassium
The Suspensions were 01 d g mzer (Silverson Machines
hydrogen phos-
Ltd., London).
topermit the particle size determination
debns, centrifugal sedimentation
as described by Allen." For cell
was carried out at speeds between 500
per volume from yeast he~ :t~to~ ~hosen weight of the compressed yeast and2500 rpm for a fixed time of 15 min and gravitational sedimentation
c~ncemrations were from 4 4 or not more than one week. Suspension wasalso done at 5C for up to 8 hr. Whole yeast suspensions were sed-
c ecked on a volume per vo-I 5% (w/v) of packed yeast. They were unentedunder gravity for period of up to 6 hr.
fiigures
II obtai
ame d for cell deb .urne basis . Th er .e was some scatter In . the Three industrial centrifuges were used: a) two intermittent-discharge
che s. The variation in phy ns IPrepared from the same weight of whole ~Sk-boWIcentrifuges for solids discharge (SAOOR 205 and SAMR 3036,
sown
v I 10 th
e fiigures which SIca . I'tires With
f II prope . yeast concentration .IS estfaha Separator AG, Oelde/Westfalia, Germany). The hydraulic bowl
oD'urne of. pa c ke d yeast. 0 ow 10 ter ms 0 f the original. . .
weight per ~penmgin both models was controlled by valves actuated by electronic
ISruptlon of the ~mers:b) a tubular bowl centrifuge (Sharples Super Centrifuge type 6P,
by using a M yeast suspension 0
Ltd C amon-Gaulin APY h n a small scale was accomplished e~~;alt Ltd., Camberley, Surrey) fitted with a2 mrn diam f~ed nozzle.
wer~ a~~~ey, SUSseX).ln all but th o~ogenizer: model 15M-8RA (APY ja k pIlot-plant centrifugation the feed suspension was held m a stirred,
of 4.9 x ~O't~~he 2feed tank and re: ~~:~expenment 4.4 liter suspension sc~e~edutank at S'C, It was pumped from the tank with a single rotor
gives a de
discrete P:~~: 1m. and at a constan/t for 29 rrnn at a valve pressure
of dIsruPtion, based on emperature of SOC. This procedure
the veast su s of the material throu :r~telO release, equivalent to four
do) : mp having a variable speed gearbox (Mono Pumps Ltd., Lon-
Iri~~alor the Westfalia SAMR 3036 centrifuge, the feed was via a cen-
"pacit p~mp With flow rate measured by a rotameter. The separation
Manlon_Gau~pe~lons on a larger s~ It e homogenizer. I Disruption of Y or the centnfuges was analyzed by the Sigma method." .s
liter, was pas:~d PY homogenizer, m~d:lwas accomplished by using a
10' kN/m' and 5o~our tImes through the 01KR3. The suspension, 50-100
h
the outflow throu The temperature inc achlOe at a pressure of 4.9 x RESULTS AND DISCUSSION
fluid was aqueous getha Iheat exchanger (~~:::e was countered by passing
Ph ySlcal
. Characteristics of Disrupted Yeast Cell Suspensions
adjusted in a jacketei ene glYcol at _ lQoC ,type J .H.E.). The cooling
~ssary the disrupted c sl\orage tank cooled b' The temperature was fmally D unnghom
. . . .
and SOC. e s were stored 0 Y ethylene glycol When nec- sUpemat ogeOlzahon of yeast the viscosity of the cell suspenSIOn and
Th vermght t . releasedant, the packed volumetric content of insoluble solids, and the
T e volumetricsolid a temperatures between
plaitedaProtem were measured and are shown in Figure I. They are
he, ratio of final to ini~~onlenl before and' .
~enerallY in the range of ~v~~Umetric solids ~~er dIsruption was checked. s a fun r
magenizerfelon 0 f the number of discrete passes through the ho-
wgu,de to the wear of the h to 0.64. It was ~ntent, M, was found to faU M.aterialfor 45% packed weight per volume whole yeast.
th~~:e;crmlned by centrif~~~gcnlzers Used O~~dthat the M value was IheYeastslu Or the "Infi'Imty " measurements was obtained by recyc l'109
sus S ~rry volume less than ,Ion, as describ~d be packed solids content 25rnin.Tberry\through the homogenizer after the four passes for a further
pensions as om I m was Y Gray t I 3 ~sedby us asv:h~e for Folin protein at this point is above that previously
ried Out 5" a lcrnative to d'l used for h,'gh ea., except
at C V'" I utio solid '
(Rheomat IS . ISCOsHy measlIrc01. n, and the ce . Sc~ncentratlon hatthe addit' maxImum releasable soluble protein.' It seems probable
with the coax'" Ont.nlves IndUMrial ~ntswere rnad~t~l~ugatlO~
was car- Wall carbohy~onal ~aterial is protein or peptide associated with the cell-
pic. The de ,~ul cyhnder system (typ r~ducts Ltd R SlOg a VISGometer The protein ra\e, whIch IS released on prolonged homogenization.
n Ity of the dISrupted yea, S,O) Which' Ulshp, Middlesex)
l '1U\PCn,io , requires a 20 ml sam- :as ses based ~e ease obeys the expected first-order kinetics for the four
n sand th .
elr Supernatants "t
T~em VisCOSit~ maximum release of 96 mglg packed yeast. I The ap-
e VIscosity of a a shear rate of 2 17 sec - I increaseS almost linearly.
parllally p un 'fi'd
It g Iyean produced by prolonged homog-
 341
CHARACTERISTIC F 01 RUPTEO BAKERS' YEAST
J40
MOSQUEIRA ET AL.
mtesareshown in Figure 6. In each in ranee the range of intervals be-
95
tween discharge and the bowl opening time wa et up by experience to
match the throughput. If the intervals between discharge were reduced
80 ""ctlyin proportion to the increased flow rate, the rise in solids content
til~ 50 mlheoverflow stream as the flow rate increased would depend only on
""wen 20 t"eflowrate with a consequent decrea e in the underflow stream. How-
ever,they were not since the interval became impractically small, par-
5 tICularly with high input solids level . Thi add to the very poor per-
~ormance of the machine at 40% (w/v) uspen ion. The underflow volume
1U7 10-6 5 ischargedis affected by the flow rate in a second way since during the
F SETTLINGRATE (rns") 10-
tmethebowl is open, the liquid volume originally present plus the in-
Ig.4. Sedimentation h
(Il) whole yeast sus . c aracteristicsof4%(w!v' . ~mmgfeed is discharged. This dilute the. underflow more at the higher
sedimentatio . ( ) ~enslOn, gravity sediment' ) whole and disrupted yeast suspensIOns:
n ; 0 disru p t e d yeast suspensio
.u" anon: '. (0) diisrupte d yeast suspension, gravity
. BOW rates.
n, centrifugal sedimentation. tu;\Similarexperiments with flow rate as the only variable and using the
(0 uarbowl and the disk centrifuges, SAOOH 205 and SAMR 3036, their
The flow was
discharg started to the machi (0 . at'mparaltve. eff ec tiveness wi respect to sohds
rveness WIth . removal to. t h e ove rfl OW
after th es. the samples were tak ne. .47 liter/min) and, after several
.f g uirnes was determined and compared with laboratory batch
lncreasm
cent
dividi e ~esumPtlOn of overflow e~:t mtervals from a time immediately
uidc ugalton. The effectiveness of the separation at the point where liq-
for av~gl tb~ volumetric solids flo;" e results have been generalized by
of the ~ead e slUdge storage space a <.rIg. 5) by the manufacturer's figure repre:erflowcommences was then found by extrapolation (Fig. 7) and
trates th e solids unsedimented Th plotting this against the percentage thereenltsIdeal behavior. For the SAOOH 205 and SAMR 3036 machines
there is ecIpartial
I retenti .
enuon of solids 0e mtercept on t h e vertical
. ..
aXIS11Ius- discha su t repre sents the mean composition of the overflow between SOlid
J
The effe:~r t~ rapid deteriorati;n i~ce the sludge space has been filled Ihevo~ges.As the bowls filled the effective sigma value decreased and
Yeastthumetncsolids content of the overflow rose steadily. For whole
on the distribuf e mterval between disc~erformance.
amined for thl Son of solids betwee arges and the bowl opening time e
oWlw fsohds content of the 6P centrifuge overflow was zero unu1 t h e
e AOOH n overflo b asulle . d
total overflow a d 205 machine Thi wand underflow was ex- Yeastthe fl ' ven at flow rates up to 147liter/min, whereas for dlsrupte
for at least threend' underflow of the rna hi IS was done by col1ecting the of E ~w rates used were from 0.45 to 5 liter/min. Data for whole
<ells
'd Ischar c me und h ..
so II s conlent of b h ges of collected r er eac set of condItIOnS Ison." . colt obtamed by similar experiments are included for compar-
ot streams. Typical ~o Ids: and measuring the packed
peratmg results at different flow

70
5-
'3
12 &J
9~
~ 50 0
l'..

~ '""i 40
~
2
0
;;;
<'
IS is 20 is

. K ~
;i'
~
;i'
'0 1 2 J 4 5
Col !f'l(' 30 p' LIOJlD FEED RATE (I mril)
Fl.' PeKen( I.;) 40 ~6 ' '
AOOH .:0,ttllln~ of ..ell dc:ttn, (from ~
vcnu the 1 "'\1'~
(
"'Iv) !rations' Distribution
Gilt" Operating
l co d"of. solids
. between overflow and underflow at variouS feed concen-
,cnled. ) lOumulah\;c. whd vo umc: of dcbn Whole yeast
of 0.23 hler; '\ rc.Uuncd d 5 collected: (.) I unsedimenlcd in an IJ)4Oi,tllbo1 the avn rftillons, and flow rates. Closed symbols represent the underflow and
0.8 htif. a, a lTluhiple of IhcumUl3livesolids pre- s
c: Sludg e space volume 0 (wI)v, Interval e
"In" 45 bow stream ,ngmalO . suspension .
concentrallons: (0 and "') 20%;
~e"ing . c; (0) 2.5 mins etwee
, Se JO n d">scharge; (0) J min, 30 sec, 20 sec, 20 sec; (e) 6 min, I
htlles: (0) \.0 se~' (",sec, c 15 sec, reading left to right on the figure. Bowl
, 7 sec.