leveking phytase introduction

Optimized Usage of Phytase

in Animal Nutrition

- Going beyond phosphorus release

- Much more release of energy

1

2

Content

Introduction of Leveking (p2-p7)

Introduction of Phytate (p8-p12)

Introduction of Leveking Phytase (p13-p19)

Experiment and Conclusion (p20-p44)

Shenzhen Leveking bio-engineering Co. Ltd. is a National High-tech enterprise,

integrating enzyme R&D, production, application and marketing. Our products

and solutions apply in food、animal nutrition, leather tech, paper-making and so

on. Leveking Animal Nutrition is a subsidiary of Leveking, focusing on enzyme

development and application in animal nutrition.

As a leading feed lipase enterprise in China, Leveking owns a strong property

rights system and obtains a number of national invention patents.

Introduction of Leveking

4 Enzyme Production Bases, ensure high quality and effective supply.

Production Base in Shenzhen

Pilot plant for high-end enzyme production.

Production Base in Ningxia

A large production base in Ningxia, North China.

Production Base in Anhui

Main production base with latest fermentation

equipment and production technology.

Production Base in Guangming

For food and feed enzyme compounding

with clean workshop of hundred thousand level.

Production Bases

Certificates

Labs and Instruments

Production Equipments

1

2

Content

Introduction of Leveking (p2-p7)

Introduction of Phytate (p8-p12)

Introduction of Leveking Phytase (p13-p19)

Experiment and Conclusion (p20-p44)

Anti-nutrient effect of phytate

2 3 4 5

Binds with protein

(Wise 1983)

Chelates with calcium and trace-elements

(Wise 1983)

Acidic pH Basic pH

1

32

56

OH

P OOH

O

H

OH

P OOH

O

H

H

OH

P OOH

O

H

O H

PO O H

O

OH

P OOH

O

H

H

O

H

P OOH

O

4

Phytate reduces the availability of protein,

calcium, trace-elements and possibly other

nutrients such as starch

pH 4.5

Disadvantage of Phytate

Phytate in the feed can prevent phosphorus absorption for monogastric

animals, which increases feed cost;

Phytate chelates with minerals in the feed so that them can not be

utilized by the animal and more inorganic minerals shall be supplemented;

Phytate will combine with amylase, protease and lipase in the feed, to

inhibit activity of the endogenous enzymes, leading to a indigestibility of

proteins and amino acids;

Phosphorus excreted to the environment leads to pollution.

0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

Average

Min

Max

Phytate levels (% IP6) in raw materials

Strong correlation between phytate breakdown and protein digestibility in vivo

Phytate Breakdown VS Digestibility

R² = 0.67

60

65

70

75

80

85

90

95

20 30 40 50 60 70 80 90 100

Tota

l AA

dig

est

ibili

ty (

%)

Phytate degradation (%)

Amerah et al., 2012

1

2

Content

Introduction of Leveking (p2-p7)

Introduction of Phytate (p8-p12)

Introduction of Leveking Phytase (p13-p19)

Experiment and Conclusion (p20-p44)

Benefits of Phytase

Improve phosphorus utilization in the feed and reduce addition of calcium

hydrogen phosphate so as to lower feed cost.

Relieve the chelation of the phytate with digestive enzymes, maximize effect of

endogenous enzymes to enhance utilization of the nutrients.

Release mineral ions like Ca, Mg, Fe and Zn and improve utilization.

Release proteins and starch which were chelated with phytate and improve

their utilization.

Reduce phosphorus excretion and improve breeding environment and reduce

pollution.

1. Leveking Phytase Overview

(1) The thermostable phytase developed by Leveking has been improved by

several times of strain improvement with genetic engineering technology. It has

increased greatly the utilization of phytase and reduced activity loss in pelleting

process.

(2) Phytase Activity Definition（GB/T18634-2009）

One unit of phytase activity, refers to the amount of phytase liberating 1μmol of

inorganic phosphate in 1 minute from sodium phytate (5.0mmol/L) at pH 5,5 and

37°C.

(3) Activity：

Granule: 5000 IU/g，10000 IU/g

2. Natural Thermostability

3. pH Stability

4. Specification

Form: pellet

Package: 25 kg/bag, paper-plastic lamination.

Storage: Shelf life of 12 months when stored at a cool, dry and ventilated

place and avoid high temperature.

Activity (IU/g) 5000 10000

Dosage for Broiler(g/T) 100 50

Dosage for Layinghen(g/T)

80-100 40-50

1

2

Content

Introduction of Leveking (p2-p7)

Introduction of Phytate (p8-p12)

Introduction of Leveking Phytase (p13-p19)

Experiment and Conclusion (p20-p44)

Design：4800 broilers of 1 day age were divided into 6 groups, 4 repeat

and 200 each repeat.

Group Diet

1 Positive Control（PC）

2 Negative Control (NC reduced by 0.13% Avail. P, 0.143% Ca, 0.03% Na)

3 NC+250 IU/kg phytase

4 NC+500 IU/kg phytase

5 NC+1000 IU/kg phytase

6 NC+2000 IU/kg phytase

Period：84 days.

Experiment I. Effect on Growth Performance of AA Broilers

Group Final Weight（g） Feed Intake（g） FCR (%)

Gender Male Female Male Female Male Female

PC 9204bc 6491bc 18292ab 13682bc 1.988bc 2.109ab

NC 8662d 6280d 17954ab 13498c 2.073a 2.150a

NC+250 IU/kg 8833d 6409c 17896b 13624c 2.026ab 2.126a

NC+500 IU/kg 9141c 6588b 17942b 13693bc 1.964c 2.079b

NC+1000 IU/kg 9375ab 6734a 18417ab 13932ab 1.965c 2.069b

NC+2000 IU/kg 9505a 6809a 18727a 14127a 1.970bc 2.075b

Experiment Results on Growth Performance of AA Broilers

0

2500

5000

7500

10000

12500

PC NC 250U/kg 500U/kg 1000U/kg 2000U/kg

Daily

Gain（

g）

Effect of Phytase on Daily Gain

Male

Female

250 IU/kg

Experiment Results on Growth Performance of AA Broilers

500 IU/kg 1000 IU/kg 2000 IU/kg

10000

12000

14000

16000

18000

20000

PC NC 250U/kg 500U/kg 1000U/kg 2000U/kg

Daily

In

take

（g）

Effect of Phytase on Daily Intake

Male

Female

Experiment Results on Growth Performance of AA Broilers

250 IU/kg 500 IU/kg 1000 IU/kg 2000 IU/kg

1.80

1.90

2.00

2.10

2.20

2.30

PC NC 250U/kg 500U/kg 1000U/kg 2000U/kg

FCR

(%

)

Effect of Phytase on FCR

Male

Female

Experiment Results on Growth Performance of AA Broilers

2000 IU/kg250 IU/kg 500 IU/kg 1000 IU/kg

Experiment II. Effect on Growth Performance and

Serum Biochemical Indicators of Broilers.

Group Diet

1Positive Control (PC, 0-3 weeks with TP 0.69%，0.45%

NPP; 3-6 weeks with TP 0.59%，0.35% NPP)

2Negative Control (NC, 0-3 weeks with TP 0.60%，0.35%

NPP; 3-6 weeks with TP 0.49%，0.25% NPP)

3 NC+500 IU/kg phytase

4 NC+2000 IU/kg phytase

5 NC+8,000 IU//kg phytase

6 NC+32,000 IU/kg phytase

Design: 576 male AA broilers of 1 day age were divided into 6

groups, 8 repeat for each group, 12 broilers each repeat. Two periods: 0-

3 weeks and 3-6 weeks.

Grouping:

(Published in No.4, Vol38, 2011, Animal Science and Veterinary Medicine In China )

Composition

1-21 day age 22-42 day age

PC LP Diet PC LP Diet

Corn 57.08 57.54 59.47 59.92

Soybean oil 0.83 0.69 2.07 1.92

Soybean meal 37.01 36.92 34.84 34.75

Fish meal (Peru) 1.00 1.00 --- ---

Methionine 0.29 0.29 0.14 0.14

Limestone 1.01 1.41 1.13 1.54

DCP 1.98 1.35 1.5 0.93

Salt 0.30 0.30 0.30 0.30

Premix 0.50 0.50 0.50 0.50

Choline chloride --- --- 0.1 0.1

Diet Composition

Table 1. Diet Composition

Nutrient

1-21 day age 22-42 day age

PC LP Diet PC LP Diet

ME（MJ/kg） 12.14 12.14 12.26 12.26

Crude protein（%） 21.50 21.50 20.00 20.00

Crude fat（%） 3.33 3.20 4.51 4.38

Ca（%） 1.00 1.00 0.90 0.90

TP（%） 0.69 0.60 0.59 0.49

NPP（%） 0.45 0.35 0.35 0.25

Ca : P 1.45 1.67 1.53 1.84

Diet Nutrition

Table 2. Diet Nutrition

Initial Weight（g）

Final Weight （g）

Average Daily Intake（g）

Average Daily Gain（g）

FCR (%)

PC 38.69± 0.34a 761.05± 22.42ab 45.06±1.34a 34.31±1.07ab 1.31±0.050a

NC 38.70± 0.45a 743.09±24.32b 43.90±1.26a 33.45±1.16b 1.32±0.042a

NC+500 IU/kg 38.15± 0.79a 762.90±41.46ab 45.06±2.09a 34.39±1.98ab 1.31±0.084a

NC+2000 IU/kg 38.42± 0.24a 775.59±13.78ab 45.20±2.58a 34.40±0.66a 1.29±0.045a

NC+8000 IU/kg 38.05± 0.60a 777.58±45.36ab 45.03±2.11a 35.09±2.16a 1.28±0.072a

NC+32000 IU/kg 38.23± 0.83a 785.03±31.33a 45.39±0.89a 35.45±1.49a 1.28±0.050a

Table 3. Effect on Growth Performance of Broilers of 1-21 day age

Experiment Results

Initial Weight（g）

Final Weight （g）

Average Daily Intake（g）

Average Daily Gain（g）

FCR (%)

PC 761.05± 22.42ab 2095.63±105.82c 113.80±5.56a 47.78±5.26c 2.33±0.20a

NC 743.09± 24.32b 2061.44±82.55c 110.73±4.72a 46.60±3.05c 2.45±0.18a

NC+500 IU/kg 762.90± 41.46a

b 2098.31±78.19c 111.92±5.44a 48.09±2.18c 2.33±0.20a

NC+2000 IU/kg 775.59± 13.78a

b 2192.75±84.74b 111.05±4.30a 52.29±4.57b 2.14±0.18b

NC+8000 IU/kg 777.58± 45.36ab

2332.94±102.41a 111.19±6.85a 57.48±4.24a 1.94±0.07c

NC+32000 IU/kg 785.03±31.33a 2344.88±81.21a 114.32±2.90a 56.90±2.90a 2.01±0.13b

Table 4. Effect on the Growth Performance of Broilers of 22-42 day age

Experiment Results

113.8 110.73 111.92 111.05 111.19 114.32

40.00

60.00

80.00

100.00

120.00

140.00

PC NC +500 +2000 +8000 +32000

Daily

In

take

（g

/d）

Graph - Table 4

Effect on Daily Intake of Broilers of 22-42 day age

Experiment Results

47.78 46.60 48.09 52.29

57.48 56.90

20.00

30.00

40.00

50.00

60.00

70.00

PC NC +500 +2000 +8000 +32000

Daily

Gain（

g/d

）

Graph - Table 4

Effect on Daily Gain of Broilers of 22-42 day age

Experiment Results

2.332.45

2.332.14

1.94 2.01

0.50

1.00

1.50

2.00

2.50

3.00

PC NC +500 +2000 +8000 +32000

FCR

(%

)

Graph - Table 4

Effect on FCR of Broilers of 22-42 day age

Experiment Results

Group Ca（mmol/L） P（mmol/L）Total Protein

（g/L）SAP（U/mL）

PC 2.89± 0.41ab 3.36±0.52a 27.02±5.40a 3.59±0.66c

NC 2.88± 0.22b 2.72±0.43c 25.78±3.01a 5.55±1.19a

NC+500 IU/kg 3.13±0.40a 3.29±0.31ab 27.03±2.51a 4.87±1.42ab

NC+2000 IU/kg 2.98±0.31ab 3.17±0.45ab 27.15±2.22a 4.60±1.66abc

NC+8000 IU/kg2.86±0.31b 3.12±0.33ab 27.30±2.10a 3.85±1.35c

NC+32000 IU/kg 2.76±0.30b 3.06±0.37ab 27.53±2.01a 3.56±0.96c

Table 5. Effect of Phytase on the SAP( serum alkaline phosphatase) Activity, Total

Protein, Calcium and Phosphorus Levels of 21-day age Broilers

Experiment Results

Ca（mmol/L）

Phosphorus（mmol/L）

Total Protein（g/L）

Alkaline phosphatase（U/mL）

PC 2.61± 0.20a 2.03±0.18bc 31.61±4.97ab 1.45±0.41b

NC 2.60±0.35a 2.00±0.32c 31.34±6.78ab 1.87±0.34a

NC+500 IU/kg 2.60±0.13a 2.11±0.22abc 33.29±6.32ab 1.68±0.73ab

NC+2000 IU/kg 2.57±0.18b 2.16±0.22ab 33.89±6.96ab 1.46±0.38b

NC+8000 IU/kg 2.57±0.21b 2.20±0.25a 34.90±5.41ab 1.43±0.33b

NC+32000 IU/kg 2.51±0.19b 2.19±0.19a 35.08±6.60a 1.45±0.23b

Table 6. Effect of Phytase on the SAP (serum alkaline phosphatase) Activity, Total

Protein, Calcium and Phosphorus Levels of 42-day age Broilers

Experiment Results

2.61 2.60 2.60 2.57 2.57 2.51

1.50

1.80

2.10

2.40

2.70

3.00

PC NC +500 +2000 +8000 +32000

Ca（

mm

Ol/

L)

Graph – Table 6

Effect on Serum Ca Level of 42-day age Broilers

Experiment Results

2.03 2.00 2.11 2.16 2.20 2.19

0.50

1.00

1.50

2.00

2.50

3.00

PC NC +500 +2000 +8000 +32000

P（

mm

ol/

L)

Graph – Table 6

Effect on Serum Phosphorus Level of 42-day age Broilers

Experiment Results

Conclusion

Supplement of phytase from 500 IU/kg increased to 2000 IU/kg can

improve daily gain and FCR further.

According to plenty of experiment data, 2000 IU/kg of Leveking

phytase can release 0.15% of available P. and 40-50kcal/kg of energy.

The accurate remaining activity of phytase can be aasayed by

pelleting recovery rate, so as to help nutritionist optimize the usage

of phytase.

Thanks for your attention!