Kolida S | Hernandez O | OHara SP| Rastall RA|

Contact: Dr Sofia Kolida | R&D Director | OptiBiotix Health PLC |Tel: ++ 44 7585001450|

BackgroundLactobacillus plantarum LPLDL

Ò is a probiotic with establishedcholesterol reducing activity in normal to mildly hypercholesterolemicadults. Probiotics can reduce cholesterol by various mechanisms:

• Adsorption to cell surface.• Integration to cell membrane.• Bile Salt Hydrolase (BSH) activity- prevents bile salt reabsorption.

• Cholesterol esterase activity-conversion to coprostanol.

Mechanisms of cholesterol reduction by Lactobacillus plantarum LPLDL®

AimsInvestigate the relevance of the above described mechanisms to thecholesterol lowering activity of LPLDL

Ò.

MethodsTo investigate the ability of LPLDL

Ò of passively removing cholesterolthrough adsorption on its cell membrane, pure cultures were carriedout in the presence and absence of cholesterol. Samples wereanalysed using scanning electron microscopy (SEM).Faecal, micro-scale, pH-controlled batch cultures (10ml) containingcholesterol-enriched growth media, were carried out in the presenceof LPLDL

Ò . Culture supernatants were obtained at 0, 10 and 24h for:• Bile Salt Hydrolase (BSH) activity determination.• Cholesterol and coprostanol concentrations were determinedusing GC-FID.

A double blind, placebo controlled, randomized study was carried outin normal to mildly hypercholesterolemic adults to confirmcholesterol reducing efficacy in vivo.

Results

Adsorption of cholesterol on LPLDLÒ cell surface: SEM of LPLDL

Ògrownin the absence (A) and presence of cholesterol (B) following washingin PBS.

A B

BSH activity in pH-controlled faecal batchcultures: significantincrease in bile salthydrolysis at 8 and 24hwith significantly higherpreference for glycolate.

0

50

100

150

200

250

300

350

Control Control LPLDL® Control LPLDL®

0h 8h 24h

Activ

ity u

nits

/ml

Glycholate Glycodeoxycholate Taurocholate Taurodeoxycholate

0

5

10

15

20

25

30

35

Control Control LPLDL® Control LPLDL®

0h 8h 24h

% c

hole

ster

ol re

mov

al

Cholesterol reducing activity in pH-controlled faecal batchcultures: significant increase in cholesterol reduction at 8 and24h in cultures where LPLDL

Òwas present.

Coprosanol formation in pH-controlled faecal batchcultures: Conversion ofcholesterol to coprostanol inthe presence of LPLDL

Ò after 8and 24h fermentation. Onceformed, coprostanol isexcreted in faeces in vivo.

0 h

8 h

24 h

Impact of LPLDL® intake on serum lipid profiles in vivo:

statistically significant reduction in systolic blood pressure, LDLand total cholesterol and an increase in HDL cholesterol innormal to mildly hypercholesterolemic adults

Study HighlightsOur findings suggest that LPLDL

Ò can mediate cholesterolreduction by three mechanisms:I. Adsorption

II. BSH activityIII. Enzymatic conversion to coprostanolThe ability of LPLDL

Ò to reduce cholesterol through severalmechanisms contributes to its enhanced efficacy in improvingserum lipid profiles in human studies.