Professional Documents
Culture Documents
5
5
5
86:2744–2750
© American Dairy Science Association, 2003.
2744
MILK FAT GLOBULE MEMBRANE FRACTIONS FROM BUTTERMILK 2745
buttermilk would lead the way to the commercialization cutoff (Synder Filtration, CA) and Supor 100, 0.1-mm
of a novel ingredient with unique functional properties. hydrophilic polyethersulfone (Pall Gelman, Ann Arbor,
The production of different buttermilk fractions by MI). Volumes between 8 and 16 L of buttermilk were
adsorption on various biosilicates has recently been re- used in each experiment. Most experiments of ultrafil-
ported (Fryksdale and Jiménez-Flores, 2000, 2001). tration and microfiltration were carried out at 50°C,
Biosilicates show a higher affinity for phospholipids but some preliminary work was also carried out at 10°C
than neutral lipids. More research needs to be done to with membranes of 250,000 and 500,000 Da. The but-
optimize the fractionation technologies and to obtain termilk retentate was circulated through the system
isolated MFGM with small amounts of contaminant by a 2-HP centrifugal feed pump (Hydracell Wanner
proteins; indeed, the isolation of MFGM and the recov- Engineering Inc., Minneapolis, MN) until reaching the
ery of phospholipids from commercial buttermilk will desired final concentration, and the temperature was
be possible only if caseins, whey proteins, lactose, and maintained by a shell and tube heat-exchange system
minerals are selectively removed. maintaining the temperature with a circulating water
Membrane filtration has been employed for bacterial bath (Neslab HTC 1200 Newington, NH). Pressure was
removal in milk (Huffman and Harper, 1999), but it controlled by valves on inlet and outlet permeate ports,
has also been used in casein enrichment, to modify the and runs were carried out at a retentate pressure of
αs/β-casein ratio of milk and for lactose separation in 1.2 MPa. Membranes were cleaned using a cycle of
whey protein isolation (Mistry and Maubois, 1993). The 0.29% nitric acid, and after rinsing, 0.029% detergent
difficulty of separation of MFGM from casein micelles (Ultrasil II, Klenzade Ecolab Inc., St. Paul, MN). Mem-
by microfiltration of buttermilk has been described by branes were considered clean when the water flux re-
Sachedva and Buchheim (1997). Because of the compa- covered its original value, and they were stored in a
rable size of those components, these authors proposed solution of 0.28% sodium bisulfate.
the removal of caseins from reconstituted buttermilk The effect of sodium citrate on the separation was
by precipitation of the caseins and subsequent filtration also evaluated by adding sodium citrate at a ratio of
of the buttermilk serum phase, rich in phospholipids. 1:4 (wt/wt, total buttermilk solids) to reconstituted but-
By microfiltration of this buttermilk whey with 0.2-μm termilk. After the addition of citrate, the buttermilk
membranes, the authors reported a recovery of 67% of was stored overnight in a cold room (6°C) before fil-
the total phospholipids, but the yields varied depending tration.
on coagulation conditions. The final buttermilk extract Diafiltration was conducted at 50°C after the concen-
obtained contained about 1.5% phospholipids and 9% tration steps by adding a volume of deionized water
protein on a dry basis (Sachedva and Buchheim, 1997). corresponding to the permeate volume collected, unless
The work presented in this paper describes a study otherwise indicated. Multiple diafiltration steps were
on the isolation of MFGM fractions of commercial but- carried out as indicated in the results. Retentate and
termilk by membrane filtration. It is demonstrated permeates were collected, analyzed for protein concen-
that, by microfiltration of buttermilk in the presence of tration and freeze-dried.
agents that dissociate casein micelles, it is possible to
obtain a concentrate with a high ratio of membrane
Characterization of the Isolates
material to skim milk-derived proteins.
The isolates were characterized for composition with
MATERIALS AND METHODS a carbon, hydrogen, and nitrogen analyzer (PE 240C
Perkin Elmer, Boston, MA, at the University of Georgia
Microfiltration Experiments Chemical Analysis Laboratories), after microwave di-
Buttermilk powder was purchased from Dairy Farm- gestion in acidic conditions. The lipid content was deter-
ers of America (Kansas City, MO). Before drying, the mined by Sohxlet analysis. The effectiveness of filtra-
buttermilk had been pasteurized at 76°C for 30 s (ac- tion was determined by analyzing the protein composi-
cording to manufacturer’s information). However, it is tion of permeates and retentates by SDS-PAGE using
not known what heat treatment the cream had under- precast gels (Bio-Rad, Hercules, CA) with 4 to 20% gra-
gone before the butter-making process; this is discussed dient. Unless otherwise indicated, fresh retentates and
below. Reconstituted buttermilk powder (8%, wt/vol, in permeates were analyzed by dissolving 1 volume of
deionized water) was separated by membrane filtration sample in 2 volumes of sample buffer containing 60
using a pilot-scale batch filtration system (NIRO filtra- mM Tris-HCl, pH 6.8, 25% glycerol, 2% SDS, 5% 2-
tion, Hudson, WI). The system was equipped with stack- mercaptoethanol, and 0.0125% bromophenol blue as
able membranes of various nominal pore size diameter suggested by the manufacturer (Bio-Rad). Freeze-dried
polyvinylnilfluoride (PVDF) 250,000 and 500,000 Da samples were dissolved directly in sample buffer, to
CONCLUSIONS
The addition of sodium citrate to buttermilk was nec-
essary to disrupt micellar caseins and allow permeation
of the skim milk-derived proteins through a 0.1-μm
filter. The process resulted in a protein isolate con-
taining a high ratio of MFGM material. Considerable
quantities of whey proteins (especially β-lactoglobulin)
were present with the MFGM material, as a conse-
quence of the processing history of the commercial prod-
uct used. We concluded that microfiltration has poten-
tial as an industrial scale means of isolating MFGM
fractions from buttermilk. To date, this is the only suc-
cessful method in extracting MFGM with very little
skim milk contaminants from a commercial buttermilk
source. More research needs to be carried out to deter-
mine the differences in products by using varying
sources of buttermilk with different processing history,
and the possible utilization of the dissociated casein
fractions present in the permeate. More understanding
of the effect occurring to the MFGM during processing
of buttermilk powders and the cream they derive from,
would further improve the quality of the ingredient that
Figure 6. Elution chromatogram of the lipid fraction injected on could be prepared by microfiltration.
silica column (Spherisorb 80, Alltech, Nicholasville, KY) A. Butter-
milk retentate with citrate added (milk fat globule membrane isolate);
B. Buttermilk retentate, no citrate added. The elution volume of ACKNOWLEDGMENTS
standard samples is indicated by arrows, from left to right, phosphati-
dyl ethanolamine (PE), phosphatidyl choline (PC), sphingomyelin This work was funded in part by Danone Vitapole,
(SM). le Plessis-Robinson, France.