WO2012050979A1 - Novel feed additive - Google Patents

Abstract

A novel animal feed additive containing L-glutamine.

Description

NOVEL FEED ADDITIVE

CROSS-REFERENCE TO RELATED APPLICATIONS

None.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not Applicable.

BACKGROUND OF THE INVENTION

This invention relates to novel feed additives containing L-glutamine.

Glutamine Supplementation in Animal Diet

L-Glutamine is the most abundant amino acid in the bloodstream and muscles, accounting for 30 to 35% of amino acids Nitrogen in the plasma and in the free amino acid pool in the body (Newsholme et al, 1985). It is the major vehicle for nitrogen transfer between tissues. Glutamine is the principal metabolic fuel for small intestine enterocytes, lymphocytes, macrophages, and fibroblasts (Cynober, 1999, Andrew and Griffith, 2002) and is considered an essential amino acid in some species under inflammatory condition such as infection and injury (Newsholme, 2001).

L-Glutamine has long been used as a dietary supplement and pharmaceutical ingredient for human health and wellness. Its benefit includes three aspects: First, L-glutamine promotes protein synthesis and inhibits protein degradation. Its supplementation is very popular in body-builders for muscle building and for energy during exercise. Second, L-glutamine aids building intestinal mucosa, maintaining gut barrier function, and decreasing intestinal permeability, which reduces intestinal infection, septic morbidity and the symptoms of Irritable Bowel Syndrome (IBS). Third, L-glutamine is an ammunonutrient that stimulates immunocyte proliferation and increases cytokine (IL-1) secretion, which boost immune response. L-glutamine is used extensively in treatment of serious illness such as aiding gastrointestinal tract function and recovery after infection, trauma, and surgery. In recent years, the beneficial effect of L-glutamine has also been demonstrated in livestock animals. For muscle growth, Yi et al (2001) reported that supplementing the standard corn-soybean mean (SBM) diet with 1% L-glutamine improved weight gain and feed efficiency of turkey poults. Kitt et al (2002) also reported that the addition of 1% L-glutamine to the diet improved the feed efficiency in weanling pigs. Bartell et al (2007) reported that 1% supplementation of L-glutamine improved chicken weight grain by 11% in 21 days. Chen et al (2009) reported that ovo injection of L-glutamine improved the pectoralis muscle mass by 25% in ducks. Wang et al (2008) reported that 1% supplementation of L-glutamine increased the body weight gain of piglet by 19%. Dai et al (2009) demonstrated that L-glutamine supplementation not only improved growth performance, but also improved meat quality and color stability of broilers under heat stress.

L-Glutamine provides as much or more energy than glucose to the digestive tract. Intestinal mucosal cells, known as the enterocytes, utilize glutamine as a major energy source (Fleming et al, 1997). It has been determined that 35% - 40% of the carbon within glutamine is utilized for ATP formation versus 7% - 10% of glucose carbon. Notably, glutamine and ketone bodies (butyrate, for example) are the main respiratory fuel in the post-absorptive small intestine (Windmueler and Spaeth, 1978). It has been estimated that the entire mucosal lining of the alimentary canal is shed approximately every 24 hours. Therefore, maintaining the intestinal mucosa itself is a major drain on the energy pools of the entire body. Repairing and building the mucosa with adequate amounts of glutamine spares precious energy pools. Increased energy production leads to improved diet/nutritional status enhancing feed efficiency and growth performance. Another benefit of raised energy is improved motility and pulsation of the intestines, contributing to efficient digestion.

L-Glutamine supplementation increased intestinal villus heights in poults (Yi et al, 2001), weanling pigs (Kitt et al, 2002; Yi et al, 2002), and broiler chicks (Bartel and Batal, 2007; Murakami et al, 2007). In experimental animals and humans, L- glutamine supplementation has been reported to stimulate gut mucosal proliferation in rats (Inoue et al, 1993). It has been observed that supplementing with 1.5% L-glutamine in total parenteral nutrition diet maintains gut integrity (Naka, 1996), which is important in preventing bacterial infections. L-Glutamine has been shown to prevent intestinal hyper-permeability and bacterial translocation in mice during an immunological challenge (Adjei et al, 1994). Under stressful conditions, intestinal permeability increases allowing bacteria to enter the bloodstream, thus causing infection (Adjei et al, 1994). L-glutamine has also been shown to decrease the incidence of infection in surgery and trauma patients (Newsholme, 2001; Medina, 2001; Adrew and Griffiths, 2002). In a more recent experiment, although 1% supplementation of L-glutamine was not able to reduce Salmonella colonization in the ceca of young broiler chicks the supplementation did stimulate the resistant against the bacterial infection (Fasina et al. 2010),. Indeed, supplementation of 1%> L-glutamine reduced the incidence of Eimeria maxz^'ma-induced lesions in the intestine of broiler chicks (Yi et al. 2005).

It is well established that L-glutamine enhances immune response. Bartel and Batal (2007) reported that supplementation of L-glutamine stimulate the humoral immune response of broilers. The birds fed diet supplemented with 1%> of L-glutamine for seven days or more had significantly higher IgA concentrations in the serum and bile than the control birds. The supplemented group also had significantly higher IgG concentration in the serum. In weaning piglets, L-glutamine supplementation effectively alleviated the immune suppression caused by lipopolysaccharide (LPS) (Jiang et al, 2009) or by Escherichia coli K88⁺ (Yi et al, 2005) challenges. The alleviation is most likely mediated by reduced secretion of IL-Ιβ. The same phenomena as also reported in broilers challenged with Salmonella Typhimurium (Fasina et al. 2010).

While L-glutamine is increasingly demonstrated to be an excellent feed additive in experiment, it is not used as a practical feed additive livestock animal yet. First, it is too costly to add 1%> of purified L-glutamine in animal feed. Second, production of purified L-glutamine needs large amount of energy and generate considerable amount of wastes including organic wastes, which makes the process environmentally not sustainable. Third, purified glutamine is difficult to supplement because it is poorly soluble in water and it is readily precipitate out from solution. To overcome these short-comings of purified L-glutamine, Chengzhi Life Science Co Ltd, the long time manufacturer of L-glutamine, has recently developed a new process technology that produced L-glutamine in a nutritional mix, BioGlutamine™, which is more suitable as feed additive.

The art described in this section is not intended to constitute an admission that any patent, publication or other information referred to herein is "prior art" with respect to this invention, unless specifically designated as such. In addition, this section should not be construed to mean that a search has been made or that no other pertinent information as defined in 37 C.F.R. § 1.56(a) exists.

BRIEF SUMMARY OF THE INVENTION

The invention provides an additive that is a better feed additive than previously available. Comparing with pure L-glutamine, BioGlutamine™ feed additive has much more nutritional components. First, it is a protein source, having over 30% of ready digestible protein (Table 1). Second, it contains other amino acids such as leucine, iso leucine, methionine, threonine, and cysteine essential amino acids (Table 2). Third, BioGlutamine™ A feed additive contains live bacteria that can serves as probiotic, which is beneficial for animal growth and health.

DETAILED DESCRIPTION OF THE INVENTION

Manufacturing Process of BioGlutamine™ Feed Additives

We have developed two manufacturing processes for BioGlutamine™ feed additive production. The first process takes the advantage of our existing glutamine process and produces BioGlutamine™ feed additives as a co-product of L-glutamine (as shown in Figure la). The co -pro duct approach minimizes the cost and wastes associated with L-glutamine and BioGlutamine™ feed additives production. However, as a co- product, BioGlutamine™ feed additives production is limited by L-glutamine co- production. In order to produce large quantities of BioGlutamine™ feed additives, a dedicated process is developed (as shown in Figure lb). Each of these processes produces a stable BioGlutamine feed additive product. Depending on the process used and processing aid employed, the L-glutmanine content in the final feed additive product varies from 8% to 36%. Table 1 examplifies two products that contains 8% or 24% L- glutamine

Process Flow Chart for BioGlutamine Production

(a)

Figure 1. Process Flow Chart for BioGlutamine Production. Two manufacturing processes for BioGlutamine™ production have been developed. The first process takes advantage of our existing glutamine process and produces BioGlutamine™ as a co- product of L-glutamine (as shown in Figure la). The co-product approach minimizes the cost and wastes associated with glutamine and BioGlutamine™ production. However, as a co-product, BioGlutamine™ production is limited by L-glutamine co -production. In order to produce large quantities of BioGlutamine™, a dedicated process is developed (as shown in Figure lb). Each of these processes produces a distinct BioGlutamine™ product. Process A produces a BioGlutamine™ that contains 8% glutamine while process B gives out a product that contains 24%> of glutamine.

Suitable raw materials include glucose, corn steep liquor and ammonia sulfate. The fermentation may be at about 32°C, with a pH of between about 6-6.5 and dissolved oxygen of about 10-13%. Fermentation is for between about 36 - 48 hours. Flocculation, precipitation, filtration, the filtrate, which is rich in protein, can be added back into the broth. The mixture can be further mixed with processing aid and dried to produce animal feed.

The anions and cations ion exchange are alternatively used to remove anions and cations, GA, soluble protein, and color from the broth. Vacuum concentration at 40-45 degree Celsius to reach 20-45% solids. The concentrate is cooled down slowed under gentle agitation. L-glutamine crystallizes while temperature drops. When temperature reaches 5 degree Celsius, the crystallization is terminated and the crystal- liquid mixture is centrifuged to obtain L-glutamine coarse crystal.

The coarse crystal is dissolved into pure water under room temperature and agitation. The coarse crystal contains colored impurities. After the crystal is dissolved, active carbon is added to remove the colored impurities under 45-50 degree Celsius. The active carbon is removed during filtration using 0.22 um filter. The decolored L-glutamine solution is recrystallized to obtain pure L-glutamine. Micro filtration may use a 0.22μ size. Vacuum concentration at 40-45 degree Celsius to reach 20-45%) solids. The concentrate is cooled down slowed under gentle agitation. L-glutamine crystallizes while temperature drops. When temperature reaches 5 degree Celsius, the crystallization is terminated and the crystal- liquid mixture is centrifuged to obtain L- glutamine coarse crystal.

After centrifugation, the L-glutamine crystal is dried to obtain final product. The mother liquor still contains 8-10% of L-glutamine, which is disposed under regular product condition. This 8-10% L-glutamine is recovered here to produce animal feed.

The purpose of a processing aid is to absorb water so L-glutamine can be absorbed onto the material and dried to proper animal feed. For this purpose, materials, such as fiber, usually used in animal feed are used. Vacuum drying, 40-50 degree Celsius, for 4-8 hours. The dried material is further milled to reach a particle size of 20-80 mesh.

Charactieristics of BioGlutamine Feed Additives

The BioGlutamine™ products have been analyzed. Table 1 is a summary of their general properties.

Table 1 : Summary of BioGlutamine™ Products

Generally, BioGlutamine B feed additive contains more nutrients (L- glutamine, glutamic acid, and protein) than BioGlutamine™ A feed additive. However, BioGlutamine™ A feed additives contain bacterial cells that could serves as probiotic function. BioGlutamine C feed additives is a liquid form, which a different processing aids were utilized.

Nutritional Analysis of a BioGlutamine™ Feed Additive

A detailed nutrient analysis, especially amino acid profile, has been performed on BioGlutamine™ A, B, and C feed additives to evaluate their nutritional value. It can be seen clearly that BioGlutamine™ A and B feed additives have very similar nutrients profile since there starting material and processing parameters are essentially similar. BioGlutamine™ B feed additives contains higher concentration of glutamine and glutamic acid than BioGlutamine™ A feed additives. BioGlutamine C feed additive is significantly different from A and B because different processing aid are utilized.

Table 2. Nutritional Profile of BioGlutamine feed additives

Digestibility of BioGlutamine Feed Additives

Amino acid digestibility of BioGlutamine™ feed additives have been tested by the Poultry Laboratory in the University of Illinois at Urbana-Champaign. The digestibility of BioGlutamine™ A and B feed additives were determined in caged cecectomized Single Comb White Leghorn roosters and TME_n in conventional (intact) roosters. Twelve roosters were fasted for 24 hours and then tube- fed (into crop) 30 g of BioGlutamine™ A or B feed additives (six roosters each). A tray was placed under each cage and all excreta were collected for 48 hours. The excreta were freeze-dried, weighed and ground and then analyzed for amino acids (for amino acid digestibility) or gross energy and nitrogen (for TME_n). True amino acid digestibility and TME_n were then calculated using amino acid and energy excretion by fasted roosters for an endogenous correction. The resulted are shown in table 3. It is very clear that BioGlutamine™ A and B feed additives are readily digested by the bird.

Table 3: Digestibility of BioGlutamine feed additives

* The feed were BioGlutamine additives mixed 1 : 1 with corn. The corn's

contribution has been calculated out in the above values. Table 4. Analytical Result of Products from Path B.

Product #3 has high L-glutamine and protein contents with low sulfate content. This is table shows that ammonium sulfate changes in the starting materials affects the output of the animal feed.

Performance of BioGlutamine A and B Feed Additives

BioGlutamine™ products would perform much better than its purified L- glutamine. The presence of other nutrients such as amino acids and metal ion further enhances the effectiveness of L-glutamine. The branched chain amino acids (BCAA), including L-Leucine (1.18), L-Isoleucine (0.60), and L- Valine (0.89), are essential for muscle growth. The sulfur containing amino acid L-Cystine and L-methioline in

BioGlutamine™ A and B are important for egg production. BioGlutamine™ A and B feed additives also contain significant amount of Lysine, Arganine, Pheloalanine, Tyrosine, Proline, and Threonine. These amino acids are all important for animal growth.

The above disclosure is intended to be illustrative and not exhaustive. This description will suggest many variations and alternatives to one of ordinary skill in this art. All these alternatives and variations are intended to be included within the scope of the claims where the term "comprising" means "including, but not limited to". Those familiar with the art may recognize other equivalents to the specific embodiments described herein which equivalents are also intended to be encompassed by the claims.

This completes the description of the preferred and alternate embodiments of the invention. Those skilled in the art may recognize other equivalents to the specific embodiment described herein which equivalents are intended to be encompassed by the claims attached hereto.

Claims

What is Claimed:

1. A process for preparing animal feed containing glutamine comprising the steps of:

(a) fermentation of a carbon source and a nitrogen source to produce L- glutamine;

(b) separating the biomass and broth produced by fermentation;

(c) using an ion exchange on the broth; and

(d) drying and granulating the solids to thereby produce an animal feed containing L-glutamine.

2. The process of Claim 1 wherein said l-glutamine content is at least about 8% by weight.

3. The process of Claim 2 wherein said animal feed includes said biomass.

4. The process of claim 1 further including a processing aid selected from the group consisting of fiber, proteins or acceptable animal feed grade ingredients.

5. An animal feed prepared by a process comprising the steps of:

(a) fermentation of a carbon source and nitrogen source to produce L- glutamine;

(b) separating the biomass and broth produced by fermentation; and

(c) using an ion exchange on the broth; and

(d) drying and granulating the solids to thereby produce an animal feed containing L-glutamine.

6. A process for preparing animal feed containing glutamine comprising the steps of:

(a) fermentation of a carbon source and a nitrogen source to produce L-glutamine containing liquid; and

(b) processing said L-glutamine containing liquid to thereby produce an animal feed containing L-glutamine.