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

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
32
56
OH
P OOH
O
H
OH
P OOH
O
H
H
OH
P OOH
O
H
O H
PO O H
O
OH
P OOH
O
H
H
O
H
P OOH
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
TotalAAdigestibility(%)
Phytate degradation (%)
Amerah et al., 2012

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

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 Laying
hen(g/T)
80-100 40-50

 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
 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
DailyGain（g）
Effect of Phytase on Daily Gain
Male
Female
250 IU/kg
500 IU/kg 1000 IU/kg 2000 IU/kg

10000
12000
14000
16000
18000
20000
DailyIntake（g）
Effect of Phytase on Daily Intake
Male
Female
250 IU/kg 500 IU/kg 1000 IU/kg 2000 IU/kg

1.80
1.90
2.00
2.10
2.20
2.30
FCR(%)
Effect of Phytase on FCR
Male
Female
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
1
Positive Control (PC, 0-3 weeks with TP 0.69%，0.45%
NPP; 3-6 weeks with TP 0.59%，0.35% NPP)
2
Negative Control (NC, 0-3 weeks with TP 0.60%，0.35%
NPP; 3-6 weeks with TP 0.49%，0.25% NPP)
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±105.82
c 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± 45.36
ab
2332.94±102.41
a 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

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
DailyIntake（g/d）
Graph - Table 4
Effect on Daily Intake of Broilers of 22-42 day age

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
DailyGain（g/d）
Graph - Table 4
Effect on Daily Gain of Broilers of 22-42 day age

2.33
2.45
2.33
2.14
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

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/kg
2.86±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

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

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（mmOl/L)
Graph – Table 6
Effect on Serum Ca Level of 42-day age Broilers

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（mmol/L)
Graph – Table 6
Effect on Serum Phosphorus Level of 42-day age Broilers

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.

Leveking Phytase Introduction

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