CN102876757B

CN102876757B - Technique for preparing (FOs) feruloyl oligosaccharides by adopting two-period-type combined regulation fermentation technology

Info

Publication number: CN102876757B
Application number: CN201210387682.7A
Authority: CN
Inventors: 顾振新; 余晓红; 尹永祺; 屠康
Original assignee: Nanjing Agricultural University
Current assignee: Nanjing Agricultural University
Priority date: 2012-10-15
Filing date: 2012-10-15
Publication date: 2014-12-10
Anticipated expiration: 2032-10-15
Also published as: CN102876757A

Abstract

The invention relates to a technique for preparing (FOs) feruloyl oligosaccharides by adopting a two-period-type combined regulation fermentation technology, belonging to the technical field of deep processing of agricultural and sideline products. The technique is characterized by comprising the following steps of: preparing a fermentation medium by adopting wheat bran as a main material, and regulating the initial pH and the initial temperature of fermentation liquor and then inoculating agrocybe aegevita and/or aureobasidium pullulans; and during the fermentation process, regulating the pH and the temperature of the fermentation liquor again, hydrolyzing wheat bran fibers by utilizing xylanase produced by microbes, so as to prepare the FOs. With the adoption of the technique, the reduction of the steps for preparing the FOs by adopting an enzymic method is facilitated; and the technique has the advantages of simple preparation process, low preparation cost and short preparation period, and can be used for improving the utilization ratio of wheat bran resources. Due to the adoption of the two-period-type combined regulation technology, the activity of the xylanase is ensured to be at the optimum level, so that the purpose of high yield of FOs is achieved. The technique is suitable for preparation of health-food-level FOs, and the yield of the FOs can reach 1123nmol/L.

Description

A kind of two-section type combined regulating fermentation technique is prepared feruloylated oligosaccharides technique

One, technical field

The present invention relates to a kind of two-section type combined regulating fermentation technique and prepare feruloylated oligosaccharides (FOs) technique, belong to processing of agriculture product technical field.

Two, background technology

Functional oligose is a kind of of saccharide compound, has the characteristics such as sweet, sticky, can breed intestinal beneficial bacterium, preventing cancer, prevention of obesity, prevent constipation etc., edible as sucrose substitute.The feruloylated oligosaccharides (Feruloyl oligosaccharides, FOs) being extensively present in grass is a kind of of functional oligose, by the sugared hydroxyl on different positions and forulic acid (Ferulic acid, FA) carboxyl esterification, is formed.FOs has asafoetide acyl group and hydrophilic oligose group; thereby have water-soluble and thermotolerance concurrently; having solved preferably water insoluble (as carotene, phenolic compound and vitamin-E etc.) that current natural antioxidants runs into and be unsuitable for hot worked thermo-sensitivity (as vitamins C etc.) two large application difficult problems, is a kind of have higher researching value and natural antioxidants of using value.FOs anti-oxidant activity is better than forulic acid and vitamins C, and mouse red blood cell haemolysis is had to stronger restraining effect; FOs has special physiologically active to human body, can significantly suppress the growth of leukemia human lymphocyte, promotes leukemia T cell gangrene, can suppress the albumen non-enzymatic glycation in human body, and the generation of prevent diabetes etc., are widely used in protective foods in recent years.FOs has huge application potential in foodstuffs industry, but FOs efficiently prepares problem, has become the bottleneck that it is widely used.

Based on FOs, there is stronger oxidation-resistance and biological activity, find the focus that its source and high efficiency preparation method have become research.At present, conventionally to take barley endosperm, barley straw, maize bud, hominy chop, wheat bran and the bagasse etc. of grass be raw material to the raw materials of FOs.The long-term wheat yield of China is up to 1.15 hundred million tons of left and right, and the wheat bran producing in the flour course of processing, at 3000～3,500 ten thousand tons, is preparation FOs important sources.

At present, the method for preparing FOs has Physical, chemical method, biological enzyme and biological fermentation process etc.The preparation method of some FOs is disclosed both at home and abroad, paper < < Heat extraction of maize fiber hemicellulose > > (Benko Z, Andersson A, Szengyel Z, et al.Applied Biochemistry and Biotechnology, 2007, 137:253-265.) with paper < < Production of feruloylated arabinoxylo-oligosaccharidesfrom maize (Zea mays) bran by microwave-assisted autohydrolysis > > (Devin J.R., GeorgeE.I.Food Chem., 2010, 119:1613-1618.) technique that hot extraction and microwave method are prepared Fos disclosed, but the drawback of physics preparation method is to need the strict treatment condition of controlling, otherwise can cause FOs to decompose.Paper < < Isolationand structural identification of diarabinosyl 8-O-4-dehydrodiferulate from maize braninsoluble fibre > > (Allerdings E., Ralph J., Schatz P.F., et al.Phytochemistry, 2003, 66:113-124.) with paper < < Extraction and partial characterization of feruloylatedglucuronoarabinoxylans from wheat bran > > (Schooneveld-Bergmans MEF, HopmanAMCP, Beldman G, et al.Carbohydrate Polymers, 1998, 35:39-47.) reported that chemical method prepares the technique of Fos, but there is chemical substance residue problem in it, and the oxyhydroxide of high density easily causes FOs degraded, productive rate declines, making product is difficult to use in food and medicine industry, and its by product easily causes environmental pollution.It is current more common method that biological enzyme is prepared FOs, have advantages of reaction temperature and, patent (publication number CN1840673A, open day on October 4th, 2006) " a kind of method of preparing feruoylated oligosaccharide by enzymolysis of wheat bran " and patent (publication number CN 101191137A are disclosed, open day on June 4th, 2008) " a kind of method of synthesizing feruloylated oligosaccharides by biological catalysis " disclosed, it prepares Fos all needs first to utilize biological enzyme from raw material, to produce insoluble dietary fibre, do like this and both increased production link, also produce a large amount of waste water, increase production cost.

If directly by the microbe application that produces zytase in FOs preparation, the process that makes microorganism enzymatic production process and enzymolysis xylan obtain FOs unites two into one, can save the separation and purification of zymin in producing and the preparation process of insoluble dietary fibre, reduce production costs, improve productive rate.Patent (publication number CN 102174616A, open day on September 7th, 2011) disclose " a kind of method of preparing feruloyl oligosaccharides by wheat bran fermentation " and paper < < chaxingu mushroom ferment prepare wheat-bran dietary fiber and feruloylated oligosaccharides and bioactivity research > > thereof (Xie Chunyan. Nanjing: Agricultural University Of Nanjing, 2010.) all utilize fermentation method to prepare FOs, but the output of FOs has much room for improvement, can not realize it and efficiently prepare.And fermentation condition can change microbial growth speed, control the generation of meta-bolites, wherein leavening temperature, pH, dissolved oxygen amount and inoculum size etc. are to affect microorganism growth and the synthetic remarkable factor of meta-bolites.The optimization of fermentation condition can significantly improve the ability of the synthetic object product of microorganism.The research that the wheat bran of take is prepared FOs as raw material, employing two-section type combined regulating temperature and pH fermentation method is not yet reported.

Three, summary of the invention

Technical problem:

The object of the invention is to provide a kind of two-section type combined regulating fermentation technique to prepare FOs technique, with the technical problem such as solve complex production process in current Fos production technique, production cost is high, Product Safety is poor and yield poorly.

Technical scheme:

Technical solution of the present invention is summarized as follows: the wheat bran of take makes fermention medium as main raw material, inoculation fermentation microorganism after the regulation and control initial pH of fermented liquid and temperature; In fermenting process, again regulate and control fermented liquid pH and temperature, utilize the xylanase hydrolysis wheat-bran fiber of microorganisms to generate FOs.

Above-mentioned a kind of two-section type combined regulating fermentation technique is prepared FOs technique, comprises the following steps:

(1) fermention medium preparation: wheat bran 60g/L, KH ₂pO ₄1g/L, MgSO ₄7H ₂o 1g/L, VB ₁o.1g/L, in sterilizing 20min O.103MPa;

(2) once regulation and control: regulating the initial pH of fermention medium is 3.O～5.0, the organism of fermentation of inoculation activation carries out liquid submerged fermentation in substratum, and leavening temperature is 30～34 ℃;

(3) secondary regulation and control: regulating fermention medium pH while fermenting to 24～36h is 5.0～7.0,25～30 ℃ of leavening temperatures, continue fermentation 72～96h.

Beneficial effect:

Compared with prior art, a kind of two-section type combined regulating fermentation technique of the present invention is prepared FOs technique and is had the following advantages:

(1) the present invention prepares FOs take wheat bran as main raw material, the fungi tea tree mushroom with edible safety and Aureobasidium pullulans as fermentation strain, improves wheat bran resource utilization and product edible safety;

(2) in the present invention, microorganism enzymatic production process and enzymolysis xylan generate FOs process and unite two into one, and save the separation and purification of zymin in producing and the preparation process of insoluble dietary fibre, simplify production technique and also reduce production costs;

(3) the present invention adopts the zymotechnique of two-section type combined regulating temperature and pH to carry out fermentation method to prepare FOs, has broken through the research and development thinking of one-part form zymotechnique regulation and control in the past, and FOs output is high.

Four, embodiment

Embodiment 1

Wheat bran, through seasoning, is crossed 60 mesh sieves after pulverizing, preparation fermention medium, and it consists of wheat bran 60g/L, KH ₂pO ₄1g/L, MgSO ₄7H ₂o 1g/L, VB ₁0.1g/L, in 0.103MPa sterilizing 20min; Regulating the initial pH of fermented liquid is 3.0, the chaxingu mushroom bacterial classification of inoculation activation, and inoculum size is 4.5%, in 32 ℃, carries out liquid submerged fermentation; After fermentation 24h, regulating fermented liquid pH is 6.0, and in 28 ℃ of continuation fermentation 72h, FOs output reaches 894nmol/L.

Embodiment 2

Fermention medium preparation is with embodiment 1.Regulating the initial pH of fermented liquid is 4.0, the Aureobasidium pullulans bacterial classification of inoculation activation, and inoculum size is 5%, in 33 ℃, carries out liquid submerged fermentation; After fermentation 24h, regulating fermented liquid pH is 6.0, and in 27 ℃ of continuation fermentation 72h, FOs output reaches 982nmol/L.

Embodiment 3

Fermention medium preparation is with embodiment 1.Regulating the initial pH of fermented liquid is 5.0, chaxingu mushroom and the Aureobasidium pullulans mixed strains of inoculation activation, and its ratio is 1: 1, inoculum size is, 6%, in 33 ℃, carry out liquid submerged fermentation; After fermentation 36h, regulating fermented liquid pH is 6.0, and in 29 ℃ of continuation fermentation 84h, FOs output reaches 1012nmol/L.

Embodiment 4

Fermention medium preparation is with embodiment 1.Regulating the initial pH of fermented liquid is 4.0, the chaxingu mushroom bacterial classification of inoculation activation, and inoculum size is 5%, in 33 ℃, carries out liquid submerged fermentation; After fermentation 36h, regulating fermented liquid pH is 6.0, and in 29 ℃ of continuation fermentation 84h, FOs output reaches 1123nmol/L.

Embodiment 5

Fermention medium preparation is with embodiment 1.Regulating the initial pH of fermented liquid is 5.0, the chaxingu mushroom bacterial classification of inoculation activation, and inoculum size is 6%, in 32 ℃, carries out liquid submerged fermentation; After fermentation 36h, regulating fermented liquid pH is 7.0, and in 28 ℃ of continuation fermentation 84h, FOs output reaches 1078nmol/L.

More than describe embodiments of the present invention in detail, but this example of just lifting for the ease of understanding should not be considered to be limitation of the scope of the invention.Equally; any person of ordinary skill in the field all can be according to the description of technical scheme of the present invention and preferred embodiment thereof; make various possible being equal to and change or replace, but all these changes or replacement all should belong to the protection domain of claim of the present invention.

Claims

1. two-section type combined regulating fermentation technique is prepared feruloylated oligosaccharides (FOs) technique, it is characterized in that:

The wheat bran of take makes fermention medium as main raw material, inoculation fermentation microorganism after the regulation and control initial pH of fermented liquid and temperature; In fermenting process, again regulate and control fermented liquid pH and temperature, utilize the xylanase hydrolysis wheat-bran fiber of microorganisms to generate FOs, comprise the following steps:

(1) fermention medium preparation: wheat bran drying or seasoning, after pulverizing, cross 20～60 mesh sieves, then press wheat bran 60g/L, KH ₂pO ₄1g/L, MgSO ₄7H ₂o1g/L, VB ₁the mass volume ratio of 0.1g/L (w/v) preparation fermention medium, in 0.103MPa sterilizing 20min;

(2) once regulation and control: regulating the initial pH of fermention medium is 3.0～5.0, the organism of fermentation of inoculation activation carries out liquid submerged fermentation in substratum, and leavening temperature is 30～34 ℃;

(3) secondary regulation and control: while fermenting to 24～36h, regulating fermention medium pH is 5.0～7.0,25～30 ℃ of leavening temperatures, continue fermentation 72～96h.

2. according to claim 1, a kind of two-section type combined regulating fermentation technique is prepared FOs technique, it is characterized in that organism of fermentation is the bacterial classification of chaxingu mushroom, Aureobasidium pullulans single culture or both mixing, and inoculum size is 4%～6%.