Feasibility Study Report on the Industrialization of Shell Glycosaminoglycan Production Increasing Agents

Feasibility Study Report on the Industrialization of Shell Glycosaminoglycan Production Increasing Agents

Project Title: Industrialization of Shell Glycosaminoglycan Resistance and Yield Increasing Agents

Name of Institution: Dalian Baoluobao Marine Life Science Research and Development Center Co., Ltd.

I. Research purpose and significance

“Shuilingling”, a disease-resistant and disease-promoting agent of chitosylamine plants, is not easy to decompose and metamorphose, and has high activity. It is used in conjunction with farmyard manure and other biological pesticides, and pollution-free green vegetables and fruits increase production by 13% to 24%. The nutrient content increased by 4% to 19% and the pesticide residue was zero. Compared with other domestic shell glycine biopharmaceutical fertilizers, the anti-disease production effect of this formulation has reached an advanced level, and it is superior to improving the quality, flavor and appearance of agricultural products. The main purpose of this project is green agriculture, which can be applied to the production of various economic crops.

The promotion and application of chitosanamine plant disease-resistance and yield-increasing agents is a vicious circle that has emerged from the large-scale application of chemical fertilizers and chemical pesticides, and it is an inevitable requirement for achieving sustainable agricultural development. China is the world’s largest fertilizer application country, and the large-scale application of chemical fertilizers has brought about an increase in production. However, the benefits of increasing chemical fertilizer production and income have dropped from 35:1 in 1978 to the current 20:1, and the use of chemical fertilizers has continued to increase. Levels will increasingly approach the limits of the physiological needs and availability of existing crops and cause soil compaction, pollution of groundwater and the atmosphere. China is also the second largest country in the world in the application of chemical pesticides. The large-scale application of chemical pesticides controls pests and diseases, but it also directly or indirectly endangers natural enemies of humans, animals, and pests, and diseases and insect pests develop resistance and become more rampant. Annual output losses caused by diseases and insect pests are as high as 35% and economic losses are several billion yuan. To curb this vicious trend of development, it is necessary to reduce the proportion of chemical fertilizers and chemical pesticides in agricultural production and adopt more biological fertilizers. The “sapling spirit” of chitosan plant, a disease-resistant and disease-increasing agent, came into being. Without the use of chemical pesticides and 1/3 less fertilizer, the yield of pollution-free green vegetables and fruits increased by 13%-24%, and the nutritional quality increased by 3%. -19%. The preservation period is extended by 1-3 times and the pesticide residue in the product is 0. Not only that, the use of "water Lingling" can also improve the soil, protect the earthworms, and greatly increase the amount of beneficial microorganisms such as Azotobacter, fiber-decomposing bacteria, lactic acid bacteria, and actinomycetes in the soil and suppress harmful molds and viruses. Vigorously promoting the use of "water Lingling" will fundamentally improve the agricultural ecological environment and create a true sense of ecological agriculture, green agriculture.

"Water Lingling" uses food-grade raw materials and is non-toxic to non-target organisms and has good environmental compatibility. The products produced by the “farmhouse fertilizer + “water spirits” + other biological pesticides” plant protection method are pollution-free green foods of Grade A and Grade AA, and are suitable for Dalian City to implement a pollution-free green food plan and the need for foreign exchange for export. The preparation is rich in raw material resources, prices are similar to organic pesticides, and the comparative benefit of farmers' inputs and outputs is 1:17. After the project reaches its capacity, the annual output will be 7 million bags (100ml/bag), benefiting 214,000 acres of farmland, producing 1.8 billion kilograms of pollution-free green vegetables annually, increasing annual net income of 270 million yuan for farmers, and more than 440 million people will eat green without pollution Vegetables account for more than 3/4 of the total population of the city.

The research and development of this project is directly related to the development of the "121" project of the Dalian Agricultural Modernization Park. It is an indispensable and basic supporting project for the realization of green ecological agriculture, and it needs the support of governments at all levels. Making the project stronger and stronger will effectively reverse the trend of over-reliance on fertilizer and chemical pesticides in agricultural production, and produce more safety, hygiene, nutrition, and the new plant protection mode of “Farmer Fertilizer + “Shuilingling + Bio-pesticides”. The high value-added pollution-free green food meets the needs of consumers at home and abroad, making Dalian Agriculture a truly “green vegetable basket”, “green fruit basket” and “green flower basket” in northern China, Russia, South Korea, Japan, and Southeast Asia. . The majority of farmers will also use ultra-efficient bio-fertilizers from the development of this project, spend less, increase production, sell more money, and embark on a new path to get rich by relying on science to get rich.

The project unit has many advantages that are easy to support, and become stronger and bigger. The unit owns the intellectual property rights of “Chitosamine Plant Disease Resistance Enhancing Agent and Its Preparation Method” and “Seaweed Extract Plant Immunoregulatory Opsonin”, and also has the technology to further develop new marine biological fertilizer such as chitosan seed coating agent. It is expected that within two to three years, marine biotechnology will be used to build a new agricultural plant protection system based on land and sea. At present, the project unit has invested 3 million yuan, has a multi-functional supercritical fluid extraction reactor for the production of key new types of marine biological medicine fertilizer, and has installed some packaging equipment. If the gap fund is in place in time, it will only take 3 months to mass produce the first batch of chitosanamine plant disease resistance and disease increasing agents. The unit also established horizontal linkages with several domestic biopesticides companies, a number of biotechnology research institutes, and several plant protection experts with practical experience in the application of biopharmaceutical fertilizers, and can introduce dozens of new organisms into Dalian in the short term. Source and mineral source pesticides, establishing a three-tier sales and service network in cities (counties) and townships to guide the application of farmers. This unit produces, sells the new product of this unit, sells the product of other manufacturers on the agency, promotes the work by trade, can completely avoid various risks, develop healthily.

Expected goals of the project

years

project

2004

2005

2006

total

progress

Pilot and trial production

Mass production

Mass production

Annual production capacity (10,000 bags)

(100ml/bag)

28

250

300

578

Net profit (ten thousand yuan)

-26.3

248

187

435

Total tax payment (10,000 yuan)

4

169

141

314

Benefit farmland (million mu)

2

17.9

21.4

41.3

Increased agricultural output (%)

12

14

16

vegetable and fruit

Product nutrient content (mg/100g)

4

6

8

Soluble total sugar Vc, dry matter, etc.

Annual net income of farmers (ten thousand yuan)

960

20227

27392

49719

Result achieved at the completion of the project

years

project

2004

2005

2006

Instructions

Increased agricultural output (%)

12

14

16

vegetable and fruit

Product nutrient content

(mg/100g)

4

6

8

Total soluble sugar, dry matter, vitamin C

Product pesticide residue

0

0

0

Execution of quality standards, adoption of relevant national industry licenses and quality certification systems adopted by enterprises

License certification

Provincial Fertilizer Promotion

Provincial Fertilizer Promotion

National Fertilizer Promotion

Quality Certification System

ISO9000

ISO14000

HACCP

ISO9000

ISO14000

HACCP

ISO9000

ISO14000

HACCP

Second, the reporting unit

1, the basic situation of the application unit

Name of Institution: Dalian Baoluobao Marine Life Science Research and Development Center Co., Ltd.;

Address: No. 1, Block B, Building 7, Jingang Enterprise Supporting Park, No. 99 Huaihe Middle Road, Dalian Economic Development Zone;

Registration date: February 10, 2004; Registered capital: 5 million yuan; Unit registration type: limited liability;

Competent department: Economic Development Bureau of Dalian Economic and Technological Development Zone.

2, unit staff and development ability discussion

The legal representative of the unit: Liu Fujing, University, 1998. Based on research on bio-products, chemical products, and side-by-side re-marine products, he successively developed the “Chitosamine Plant Disease Resistant and Yield-increasing Agents and Its Preparation Methods” and the “Algae Extract Plants Immunopotentiated Opsonin And its manufacturing method, "Huozhimin health wine and its manufacturing method", "seaweed health wine and its manufacturing method", reported or obtained a national invention patent.

The total number of personnel in this unit is 12 people, including 8 persons with a college degree or above; 4 are the main management personnel, and the cultural level is above the college level. The age structure of this unit is young, with an average age of 32 years. The technical development personnel of this unit accounted for 50%, production personnel accounted for 33%, and sales personnel accounted for 33%.

The new product development capability of the unit: In 2005, the amount of technological development investment was 1.85 million yuan, accounting for more than 37% of the company's annual revenue. The research and development team of this unit currently focuses on the impact of marine biologically active substances on agriculture and the design of marine drug fertilizer production devices. The company has developed the Shell Glycosamine Plant Disease Resistance Enhancer Series listed as the National Spark Program Project and the Dalian Science and Technology Plan Project. It is developing an immunosuppressive opsonization factor for marine algae plant and will eventually build a new type of marine bio-medicine fertilizer. The plant protection system contributes to the realization of sustainable green agriculture in Dalian as soon as possible.

Zhang Xuecheng, technical director of the seaweed plant's immunopotentiating opsonization project, and a doctoral supervisor at Ocean University of China, are very successful in marine biotechnology such as microalgae polysaccharide extraction. Yang Kaihai, professor of Biology Department of Liaoning Normal University, has made many achievements in marine drug research and development. The above-mentioned two project technical leaders and the unit established a project contract responsibility system, and the two responsible for organizing technical forces collaborative research.

3, the unit financial status

As of December 31, 2005, the total assets of the unit was RMB 7.49 million, the total liabilities were RMB 330,000, the fixed assets were RMB 4.68 million, the sales revenue of products was RMB 2.65 million, the non-operating income was RMB 1.4 million, and the net profit was RMB 2.57 million. The current ratio was 8.46, quick ratio 6.45, return on total assets 34%, return on net assets 34%, accounts receivable turnover rate 80%. As a result, the unit has the ability to continue to promote R&D, production and operation.

4, the unit management status

The unit has established and improved various rules and regulations. In 2006, we will further strengthen the management of modern enterprises and pass the ISO9000, ISO14000 and HACCP certifications to meet international standards. The current products of this unit won the Gold Medal of China Food Fair and the honorary title of Liaoning Famous Food.

5, unit development ideas

The unit plans to take research and development of new ultra-efficient marine bio-medicine fertilizers under the support of the Economic Development Bureau of the Development Zone, and strives to establish a collection of science, industry and trade in the implementation period of 2006-2008. Modern marine bio-pesticide companies. In the near future, on the basis of the anti-disease and yield-increasing agents of chitosan plants, new products of seaweed extracts will be put on the market in batches. During the implementation period, we strived for the last project, developed a series of three products, declared two patents for inventions, and established a new marine biopharmaceutical plant protection system through three international certifications, contributing to the modernization of agriculture in Dalian. .

III. Analysis of Technical Feasibility and Maturity of the Project

(I) Technical innovation of the project

Chitosan oligosaccharide is an oligomeric substance obtained by bio-enzymatic degradation of chitosan extracted from sea crab and shrimp shells. It has multifunctional groups, multiple variable coordination numbers, and strange crystal forms. The physiological and biochemical reactions that produce the most complex life activities, coupled with its hydrophilicity, reproducibility and complete degradability, indicate that it is a type of resource-rich, excellent performance, can fully participate in the ecological system cycle, and has sustainable agriculture. Features important resources. The international academic community recognizes that the shell glycosaminophen biopharmaceutical fertilizer is a large class of new agricultural production materials whose mechanism of action is completely different from traditional medicinal fertilizers and is the leading product for the development of green agriculture in the 21st century. At present, in Japan, the United States, Europe and other developed countries, shell sugar amine bio-fertilizer has been promoted. In China, there have also been OS-Stelen, phytoalexin, Zhibaoling, Liaoshang 1, Apollo, Dopu, and other chitosan biopharmaceuticals, which are improving the soil, improving the disease resistance of crops, and promoting crops. Growth and the preservation of agricultural products have played an active role.

(II) Project Innovation

The molecular optimization design of this project is based on the principle that chitosan reacts with acid to form salts and thus enhances the ability of citric acid to dissolve. Special nutrients such as chitosan and food grade citric acid with a degree of deacetylation of more than 93% and a molecular weight of <3000 are used. , Vitamin C and other food additives, in a certain structure-activity relationship, the shell glycosulphonate (no domestic and foreign shell sugar citrate this new shell sugar amine carboxyl derivatives and its mode of action, The mechanism of action was reported, and this solution was used to develop an ultra-efficient shell glycine plant anti-disease and disease-increasing agent “water spirit”.

Chitosan citrate molecular formula:

(C6H11NO4)2C6H8O7]n

Chemical Structure:

The excellent effect of the chitosanamine plant disease-resistant and yield-increasing agent “Shuilingling” is mainly reflected in the fact that the chitosan is not easy to decompose and deteriorate, and has strong affinity with the body tissues and active expression. Citric acid is the most widely distributed organic acid in vegetables and fruits. The chemical composition belongs to alpha-hydroxy acid (AHA) with a molecular weight of 192.13. It has certain antibacterial and antioxidative effects. Compared with many commonly used acidic solutions, citric acid has the strongest dissolving power, the strongest anti-decomposition ability and the best stability in the preservation of the shell sugar amine activity. According to actual observations, under the same pH conditions, the solution containing 1.2% of shell sugar amine dissolved in citric acid for 2 years had a content of shell amine of > 1%, which was dissolved by the most commonly used dilute acetic acid, namely acetic acid. The shell glycosamine content was 1.21% solution, and there was a visually recognizable significant decomposition in only half a month, and the shell sugar glycoside content was <0.03% in 14 months. The molecular weight of citric acid is very small, and it can quickly penetrate into plant pores and cell gaps by virtue of its natural affinity with plants, which can speed up plant cell regeneration. Reaction with chitosan results in the formation of a carboxylated derivative of chitosan citrate, which protects the active group NH2 and enhances the hydrophilicity of chitosan, making it easier for chitosans with a molecular weight of <3000 to preserve it. Activity, through the environmental phase in a short period of time, infiltrate the plant's biological movement phase, biodynamic phase, stimulate the development of crop cell tissue, absorb more shell sugar amine active substances into the plant, promote its conduction, distribution, accumulation, Metabolism, thereby exerting more and more physiological and biochemical effects. Shell sugar amine plant disease resistance increase agent is also rich in vitamin C, its molecular weight of 216.12, is vegetables, fruits and other crops must be stored material. In the phase of storage and environment, vitamin C greatly improves the anti-decomposition ability of shell sugar amines that can easily decompose and metamorphose in case of light, water, air, and heat, and eliminates the common decomposing and metamorphic conditions. The phenomenon that the hand has already expired, the effective shelf life has been extended from ten days to six months to more than one year. In the environmental phase-biological movement phase, vitamin C can not only protect the shell sugar amine from decomposition and deterioration, but also can be absorbed by plants as soon as possible, and enhance the ability of plants to absorb active substances such as shell glycine. In the harvesting stage of vegetables, fruits and other crops, the vitamin C enriched in the interior of the vegetables and the semi-permeable membrane of chitosan formed on the surface of the vegetable can delay the physiological aging of the vegetables and achieve the preservation purpose.

The technical innovation of this project has brought about significant changes in the product performance and use effect of chitosan biopharmaceutical fertilizer: it can (1) significantly enhance plant resistance. A sufficient number of highly active shell sugar amines rapidly enter the plants, inducing the plants to produce resistant proteins such as phytoalexin, chitinase, chitosanase and β-1,3-glucanase, effectively controlling vegetables and fruits Flowers are caused by fungi, bacteria, and viruses. At the same time, it can also produce more phytoalexins, lignin and phenolic compounds when needed to resist pests. Promotes plant growth and improves quality. A sufficient number of highly active shell sugar amines quickly enter the plant body, fully activate the physiological and biochemical functions of plant growth and development, increase the bud rate, emergence rate, promote the development of plant roots, stems and leaves prosperous, enhance the absorption and utilization of fertilizer in soil, increase Thick and enlarged leaf area, enhance plant photosynthesis, promote the transformation and accumulation of nutrients, thereby increasing yield and quality. These obvious changes were demonstrated in the relevant scientific experiments of the Dalian Academy of Agricultural Sciences in 2002. The experiment used a chitosan plant anti-disease-increasing agent “Shuilingling” to set 3 treatments of 9 spots on cucumber, grape and marigold varieties. No. 1 is biological pesticides, No. 2 is "water Lingling" 800 times liquid plus biological pesticides, and No. 3 is ordinary production pesticides. The design uses three replicates each, and each plot uses the same amount of farmyard fertilizer. Cucumber is sprayed once every 7 days. “Shuilingling”, grape, and marigold are sprayed once every 10 days. As a result, farmer fertilizer + “shuilingling” + biological pesticide No. 2 test group performs at all stages of its growth. A significant advantage.

City Agricultural Research Institute field trials

1, cucumber test

The Effect of 1-1 "Water Lingling" on Main Agronomic Characters of Cucumber

"Shuilingling" had different degrees of influence on cucumber flowering period, first female flowering stage, plant height, stem diameter and other agronomic traits. Treating cucumber with an appropriate concentration of "water Lingling" plus biological pesticides is mainly characterized by the early flowering stage, vigorous growth of plants, and low first-fruiting stage. The flowering period 2 is May 17th, while the 1st and 3rd (control) are both May 19th and the 2nd is 2 days ahead of schedule. The first female flowering place 2 is 2.80, and the 1st is 3.0. No. 3 (control) was 2.97, and No. 2 was the lowest; plant height No. 1 was 70.1cm, No. 2 was 78.0cm, No. 3 (control) was 76.3cm, No. 2 was 2.2% higher than No. 3 (control), No. 1 It is 8.1% lower than No. 3; the thickest No. 2 stem is 0.91cm, followed by No. 3 (control) 0.83cm and minimum No. 1 0.79cm.

Cucumber agronomic traits questionnaire

deal with

project

number 1

(farmhouse fertilizer, biological pesticide)

number 2

(farm fertilizer, "water spirit", biological pesticide)

No. 3 (control)

(farmhouse fertilizer, chemical pesticides)

The first flower opening date (day/month)

19/5

17/5

19/5

More than CK days

0

-2

0

First female flower

3.0

2.80

2.97

More than CK%

1.0

-6.1

0

Height (cm)

70.1

78.0

76.3

More than CK%

-8.1

2.2

0

Stem thick (cm)

0.79

0.91

0.83

More than CK%

-4.8

9.6

0

The Effect of 1-2 Water Lingling on Disease Resistance of Cucumber

The cucumbers treated with “Shuilingling” had a low degree of susceptibility, and No. 1 was used as a control to investigate the susceptibility of downy mildew and powdery mildew at the onset of disease. Each treatment investigated 50 leaves. The results showed that under the same cultivation and management conditions, the three treatments had different degrees of susceptibility to downy mildew and powdery mildew. Downy mildew: No. 1 diseased leaf rate was 12.0%, No. 2 was 6.0%, No. 3 (control) was 10.0%, Disease index No. 1 was 8.0%, No. 2 was 4.0%, and No. 3 (control) was 6.7%. Powdery mildew: No. 1 diseased leaf was 8.0%, No. 2 was 2.0%, No. 3 (control) was 6.0%, Disease index No. 1 was 4.7%, No. 2 was 3.0%, No. 3 (control) 4.0%. The above investigation results showed that only using biological pesticides to prevent and treat diseases, the effect was not as good as that of ordinary pesticides, but cucumbers treated with “Shui Ling Ling” plus biological pesticides had better disease resistance than ordinary pesticides.

Cucumber field disease situation questionnaire

project

deal with

Downy mildew

powdery mildew

blade

Number

Leaf disease progression

disease

leaf

rate

Disease index

Number of blades

Leaf disease progression

disease

leaf

rate

Disease index

0

1

2

3

0

1

2

3

number 1

50

44

2

2

2

12.0

8.0

50

46

2

1

1

8.0

4.7

number 2

50

47

2

1

0

6.0

4.0

50

48

1

1

0

2.0

3.0

No. 3 (control)

50

45

2

1

2

10.0

6.7

50

47

1

1

1

6.0

4.0

The Effect of 1-3 "Water Lingling" on the Yield of Cucumber

The harvesting period of cucumbers treated by “Shui Ling Ling” was advanced. Harvesting period 1 was June 7th, No. 2 was June 3rd, No. 3 (control) was June 5th, No. 2 was higher than No. 3 ( Controls) 2 days in advance; the number of single plants growing on the number of melons was 9.8 on the 2nd, 3 on the 3rd (control), 9.0 on the 1st, and 8.6 on the 1st; the average single melon weight was 156.7g on the 1st, and on the 2nd 162.1 g, No. 3 (control) was 155.0 g; plot production No. 2 was 31.6 kg, 13.3% higher than No. 3 (control) 27.9 kg, and No. 1 was 26.8 kg, 3.9% lower than No. 3.

Cucumber Production Survey

deal with

project

number 1

number 2

No. 3 (control)

Harvest start date (day/month)

7/6

3/6

5/6

More than CK days

+2

-2

0

Number of single plants

8.6

9.8

9.0

More than CD%

-4.6

8.8

0

Single fruit weight(g)

156.7

162.1

155.0

More than CD%

1.1

4.6

0

Community output (kg)

26.8

31.6

27.9

Converting the output per mu (kg/mu)

3,573.5

4,213.5

3,720.2

More than CD%

-3.9

13.3

0

The Effect of 1-4 "Water Lingling" on the Quality of Cucumber

According to laboratory analysis results, soluble sugar, dry matter, and vitamin C content of cucumber treated with “Shuilingling”

All were higher than the control, in which soluble total sugar No. 2 was 2.10% and No. 1 was 2.08%, which was 8.8 and 7.7% higher than that of No. 3 (control) 1.93%; dry matter No. 2 was 4.91%, and No. 3 (control) High 4.61%, No. 1 is 4.67%, lower than No. 3; Vitamin C content No. 2 is 10.34mg/100g, which is 19.95% higher than No. 3 (control) 8.62mg/100g; No. 1 is 9.19mg/100g; The number is 6.6% higher.

Cucumber Quality Test Analysis Table

deal with

project

number 1

number 2

No. 3 (control)

Total soluble sugar (%)

2.08

2.10

1.93

More than CK%

7.7

8.8

0

Dry matter (%)

4.67