superdosing of phytases · ip6 ip5 ip4 ip3 t) +500 ftu +1500 ftu 32% point in time measure of...

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Superdosing of

phytases

Mike Bedford Research Director, AB Vista Feed Ingredients

Marlborough, UK

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Traditional dosing of phytases –

matrix driven

y = 0.0562ln(x) - 0.1986 R² = 0.9999

0

0.02

0.04

0.06

0.08

0.1

0.12

0.14

0.16

0.18

0.2

0 200 400 600 800 1000 1200

AvP

% m

atri

x

FTU/kg feed

P matrix vs dose for a "0.15" phytase

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Quantum® Blue ‘super-dosing’ – 4 Point

Improvement in FCR 4 points in FCR currently worth

$7 per tonne of feed

ab

ab

bc

a

b

bc

c

1.46

1.48

1.5

1.52

1.54

1.56

1.58

1.6

1.62

1.64

PC PC+DCP PC+500 QB NC NC + 500 QB NC + 1000 QB NC + 1500 QB

Figure 3. Composite analysis of 6 trials: Body weight corrected FCR of broilers from d 0 to 35/42 (n = 35)

y = -0.0255x + 1.638 R2 = 0.9538

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QUANTUM SUPERDOSE -TRIAL SUMMARY – ADG

(FROM WEANING THROUGH TO 21/35 DAYS POST-WEANING)

80

85

90

95

100

105

110

115

120

125

130

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39

+6.26% ADG IMPROVEMENT 80% POSITIVE I.E. > 100% OF CONTROL

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QUANTUM SUPERDOSE - TRIAL SUMMARY – FCR

(FROM WEANING THROUGH TO 21/35 DAYS POST-WEANING)

80

85

90

95

100

105

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39

+3.80% FCR IMPROVEMENT 88% POSITIVE I.E. < 100% OF CONTROL

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Why ………….

1. Does superdosing work? 1. IP6 destruction

2. IP4 and 3 destruction

3. Inositol provision

2. Do we have to have high P 1. Superdosing makes animals grow faster – higher P RQ?

2. Rephosphorylation of inositol

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Activity at low substrate: In the gut, not all phytate is

soluble and susceptible to the enzyme

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

No phytase Phytase

Ph

ytat

e (

IP6

) co

nce

ntr

atio

n -

%

Soluble Phytate

Unsoluble Phytate

Leslie, 2006

Insoluble Phytate

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Pepsin inhibited at pH 2.5 by very

little phytate

100 uM

Yu et al 2012

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Protein – phytate precipitation

depends upon pH and protein

0

20

40

60

80

100

120

Casein Corn Canola meal Rice pollards SFM SBM

Pe

rce

nt

solu

ble

pro

tein

pH 2 Protein

pH 3 Protein

Kies et al 2006

www.abvista.com June 11, 2008

0

20

40

60

80

100

120


Pe

rce

nt

solu

ble

pro

tein

pH 2 Protein

pH 2 Pr+ Phyt

pH 3 Protein

pH 3 Pr+ Phyt



www.abvista.com June 11, 2008

0

20

40

60

80

100

120


Pe

rce

nt

solu

ble

pro

tein

pH 2 Protein

pH 2 Pr+ Phyt

pH 2 P+P+P

pH 3 Protein

pH 3 Pr+ Phyt

pH 3 P+P+P



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IP6 is not the only problem Phytase has to get rid of IP5IP2 as

well

Yu et al 2012

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IP6 is bad, but IP4 is not much better

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Supplementation with a novel C. braakii phytase:

Results in a ‘pool’ of IP4

Pontoppidan et al., 2012

No phytase Phytase (500 FTU)

Pontopiddan et al 2012. 25kg pigs fed 0 or 500 FTU HiPhos and after starving for 24hrs then fed treats for 30 mins, slaughtered 1 and 2 hours post feed. A=control, B =500FTU/kg

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If it is present and soluble in the stomach, it

will ppt with something in the SI

Schlemmer 2001

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IP4 and IP3 are not innocuous

Ester Nutrient R P value n

Ileal AME, kcal -0.849 <0.0001 40

Ileal DM digestibility -0.848 <0.0001 40

Ileal N digestibility -0.693 <0.0001 40

Ileal Na digestibility -0.675 <0.0001 40

Ileal Mg digestibility -0.688 <0.0001 40

Ileal Fe digestibility -0.606 <0.0001 40

Ileal iP3

Ileal iP4

Beeson et al, 2015 submitted

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Supplementation with a novel E. coli phytase:

Superdosing results in near complete destruction of phytate (IP6) and reduced

concentrations of IP4 and IP3

Walk et al., 2013 (in press)

0

0.5

1

1.5

2

2.5

3

3.5

IP6 IP5 IP4 IP3

(µm

ol/

g d

ry w

eig

ht)

PC NC +500 FTU +1500 FTU

Point in time measure of gizzard phytate of 21-day-old broilers

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0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

IP6 IP5 IP4 IP3

(µm

ol/

g d

ry w

eig

ht)

+500 FTU +1500 FTU

32%

Point in time measure of gizzard phytate of 21-day-old broilers

Supplementation with a novel E. coli phytase:



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Mansbridge et al., 2013 (Harper Adams University)

0

1000

2000

3000

4000

5000

6000

7000

PC +P +125 +500 +1000 +2000 +8000 +32000

Ph

yta

te e

ste

rs (

nm

ol/

g d

ry w

eig

ht)

Ileal phytate ester and inositol concentration of piglets 3 weeks post-

weaning

IP3 IP4 IP5 Inositol

Supplementation with a novel E. coli pytase:



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• Inositol in Chick Nutrition, Hegsted et al, 1941. Exp Biol

Med 1941 47:376-377

• Growth responses varying from 18 to 52 g in 4

weeks have been obtained upon the addition of

inositol† to several different simplified chick rations

(Table I).

Increased phytate hydrolysis may

increase Inositol production

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Inositol Provision

Zyla et al 2004

Myo-inositol None 0.10% p value

Intake 838 861 NS

Gain 537 579 0.0199

FCR 1.57 1.47 0.0573

Toe ash % 11.9 12.2 NS

P retention 56 50 0.0015

Ca retention 61 59 NS

Broilers, 1-21d post hatch, main effects on 0.65/0.27 and 0.8/0.47 Ca AvP diets

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Extra-phosphoric effects of phytase


Broiler gizzard phytate, phytate ester and inositol concentration (d21)

Superdosing Quantum Blue decreased phytate and increased inositol concentration

Part of the superdosing response may be associated with inositol provision as well as phytate destruction

ab

a

c

b

d d d

a a

b

a

b b b c c a

bc ab c c

c c

bc bc

b

a a

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

PC +DCP +500 QB NC +500 QB +1000 QB +1500 QB

µm

ol/

g d

ry w

eigh

t

IP6 IP5 IP4 IP3 Inositol

Item,

µmol/g

BW gain,

kg P = FCR P =

Tibia

ash, g P =

Phytate -0.37 *** 0.46 *** -0.21 NS

Inositol 0.46 *** -0.57 *** 0.19 NS

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Inositol interacts with phytase

It is likely part of the superdosing effect

• FCR • Inositol* Phytase interaction p<0.0143

• LSD = 0.017

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Inositol as a source of intracellular

phytate (IP6)?

0

2

4

6

8

10

12

14

16

18

20

Liver Brain Kidney Lung HumanRBC

Humanserum

Rat tissue concentration of IP6, µM

Letcher et al., 2008

Phytate (IP6) is present in tissues at various concentrations

1. Antioxidant

2. De-phosphorylated for cell signaling, Ca metabolism and inositol

production

Inositol:

1. Mammalian cell cultures

require inositol to grow

2. IP6 can be made in the

body from inositol

3. Phytate destruction may increase the

requirement for P in the

body?

25

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Kinetics differ between phytases

0

0.5

1

1.5

2

2.5

3

0 5000 10000 15000 20000

Ph

ytat

e P

re

leas

ed

(g/

kg d

iet)

Dosage of phytase (FTU/kg)

P lycii A niger E coli Quantum Blue

Red line represents 85% phytate hydrolysis, differences between phytases driven largely by pH profiles and kM

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0

0.1

0.2

0.3

0.4

0.5

0.6

0 20 40 60 80

Ph

osp

hate

/ m

M (

ad

juste

d w

ith

Meg

azym

e v

alu

es)

Time / minutes

Evolved E coli

E coli WT 1

E coli WT 2

Apsergillus

Phytases differ in appetite for lower

IP esters

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Equilibria to consider

Dietary Phytate

Nuisance factor

Insoluble Phytate

Substrate kM ???

Soluble Phytate IP3 and lower

Critical phytate concentration

High kM

Low kM

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

0 5 10 15 20 25

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• Superdosing is NOT simply using a higher

dose with a matrix .. It is

1) Using a dose >=0.2% AvP matrix but applying only

0.15% matrix max

2) Using a phytase which has ability to

1) Clear all IP6

2) Clear all IP4

3) Produce inositol

3) Ranking of phytases in this application is

different compared to that of a 0.13 matrix

Conclusions

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Beaulac et al 2015 in press

Saskatchewan data with Quantum Blue

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Beaulac et al 2015 in press

superdosing of phytases · ip6 ip5 ip4 ip3 t) +500 ftu +1500 ftu 32% point in time measure of...

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