CN104366615A - Passion fruit juice and preparation method of passion fruit juice - Google Patents

Abstract

The invention provides a preparation method of passion fruit juice. The preparation method of the passion fruit juice comprises the following steps: crushing clearly washed passion fruits, adding water and esterase for fermenting, heating, cooling and filtering. In addition, the invention also provides the passion fruit juice, the esterase and the like prepared by adopting the methods.

Description

Passion fruit fruit juice and processing method thereof

Technical field

The invention belongs to fruit juice production technical field, specifically, the esterase that the present invention relates to passion fruit fruit juice, its processing method and wherein use.

Background technology

Passion fruit originates in Nan Meizhou, also known as passionflower, gains the name because its pulp can give out the strong fragrance of tens kinds of fruit such as banana, pineapple, strawberry, lichee, lemon, mango, smoked plum.Passion fruit contains abundant protein, fat, also has multiple compounds and 17 seed amino acids such as sugar, vitamin and phosphorus, calcium, iron, potassium, nutritious.The pulp of passion fruit can be processed into fruit drink, especially can be deployed into compound juice beverage with other fruit juice (fruit juice as high in sugar content).But, mainly due to the interference of passion fruit pericarp and seed, especially its seed oil content reaches 25%, when extraction passion fruit fruit juice, usually remove seed with being careful or taking the techniques such as squeezing, whipping, and do not use sharp sword to shred, cause grease to enter fruit juice to prevent because seed is broken, affect local flavor, and grease can float, not only affect the visual effect of beverage, and be easily oxidized, produce peculiar smell.

Such as, No. 93106822nd, Chinese patent application discloses a kind of concentrated natural passionflower juice, its processing method comprise choosing fruit, rinsing, peeling, stoning, pulp starch-splitting, squeeze the juice, etc. process.

No. 93109135th, Chinese patent application discloses a kind of passionflower full fruit pulp and juice, comprise fresh fruit cleaning in its processing, soak, pericarp is separated with pulp and seed, pericarp removes exocarp after boiling, then adds pulp, and fruit particle worn into by also glue of together pulling an oar.

No. 200410022600th, Chinese patent application discloses a kind of technique taking passionflower as raw material and make fruit juice fumet, comprise passionflower fresh fruit selected, clean, squeeze, pull an oar, filter, the operation such as film is concentrated, freeze drying, packaging.

No. 200610019166th, Chinese patent application discloses a kind of manufacture method of passiflora edulis fruit juice beverage, comprises pulp and is separated rear making beating with pericarp, wherein by the beater processing that pulp is equipped with neoprene leaf with brush wiping Pi Jihe, prevents seed broken thus.

No. 201210354143rd, Chinese patent application discloses a kind of preparation method of passion fruit pulp juice beverage, comprises and being cut open by fruit, dig out pulp, carries out the separation of seed meat, is then broken into granular by isolated pulp, boils after stirring.

In the technique of prior art, also pectase can be added to decompose colloid in pulp to improve crushing juice rate, such as, No. 201310386824th, Chinese patent application discloses a kind of preparation method of passiflora edulis drink, comprise and the pericarp of passion fruit and band seed pulp are separated, band seed pulp add stir in warm water after remove seed, put into this warm water after pericarp stripping and slicing and pull an oar, then add pectinase enzymatic hydrolysis.

But, through studying for a long period of time and some fortune, the present inventor have been surprisingly found that, by adding a certain amount of esterase, especially the esterase of the present inventor's transformation, to shredding passion fruit (more seed fragmentation must be had) whole fruit fermentation process with sharp sword, the impact of grease in seed can be eliminated, both layering can not be there is, also peculiar smell can not be produced in shelf-life endometamorphism, and fruit juice color is more bright-coloured, and on the reservation former dulcet basis of passion fruit, also add trickle peanut fragrant, further increase the fragrance level of passion fruit fruit juice, on taste except the sour-sweet taste of former passion fruit fruit juice, also add trickle tannin mouthfeel, further increase the taste level of passion fruit fruit juice.

Even to need on impeller coated neoprene etc. to subtract oligospermous fragmentation during the making beating of prior art, therewith remarkable unlike, the present invention can adopt sharp sword to pulverize (e.g., chopping), without the need to being limited by the fragmentation preventing seed.So preferably in the method for first aspect present invention, in step (1), pulverizing is pulverized with sharp sword, pulverizes with the pulverizing of blade or cutting tool as used.In the specific embodiment of the present invention, sharp sword is cutter.The pulverizing that the cost of the existing routine feasible easy to use of cutter is low or cutting tool, be also convenient to automation.

Preferably in the method for first aspect present invention, the weight ratio of passion fruit in the water in step (2) and step (1) (i.e. step (1) obtain crushed products) can be 0.8 ~ 2: 0.5 ~ 1.5, be preferably 1 ~ 1.5: 0.8 ~ 1.2, be more preferably 1.1 ~ 1.3: 0.9 ~ 1.1.Most preferably be 1.2: 1.

Preferably in the method for first aspect present invention, the amino acid sequence that the esterase in step (2) has:

(1) as shown in SEQ ID NO:2;

(2) amino acid sequence of reservation esterase active amino acid sequence SEQ ID NO:2 being increased, lack and/or replace one or several amino acid residue and obtain.

According to the amino acid sequence as shown in SEQ ID NO:2 of the present invention, by handbooks such as " Molecular Cloning: A Laboratory guides ", the gene order being cloned in expression vector and/or host cell that those skilled in the art are changed by synthesis is expressed, can purifying obtain to amino acid sequence SEQ ID NO:2 increase, lack and/or replace one or several (preferably five and following, more preferably three and following) amino acid residue and the amino acid sequence of reservation esterase active that obtains.In the specific embodiment of the present invention, the amino acid sequence of esterase is as shown in SEQ ID NO:2.

In addition, present invention provides the nucleic acid (gene) of this esterase of coding.This nucleic acid, by conventional DNA recombinant technique, can import expression vector (e.g., plasmid) and/or host cell (e.g., yeast), expresses and obtains this esterase.In the specific embodiment of the present invention, the nucleotide sequence of the nucleic acid (gene) of coding esterase is as shown in SEQ ID NO:1.

The temperature and time of fermentation can be determined according to used esterase.Preferably in the method for first aspect present invention, in step (2), the temperature of fermentation is 25 ~ 55 DEG C, is preferably 30 ~ 40 DEG C, is more preferably 32 ~ 37 DEG C, as 35 DEG C.Also preferred in the method for first aspect present invention, in step (2), the time of fermentation is 5 ~ 72 hours, is preferably 12 ~ 48 hours, is more preferably 20 ~ 32 hours, as 24 hours.

The tunning that step (2) obtains needs sterilizing, can sterilizing after Separation of Solid and Liquid, also can first sterilizing Separation of Solid and Liquid again.Because filter residue can directly or after sugaring be used as cake shape food, therefore in order to reduce the complexity of sterilizing, the present invention adopts the mode of first sterilizing Separation of Solid and Liquid again.Preferably in the method for first aspect present invention, in step (3), the temperature reached that heats up is 80 ~ 95 DEG C, preferably 82 ~ 90 DEG C, as 85 DEG C.Also preferred in the method for first aspect present invention, in step (3), the temperature that keeping heats up reaches 1 ~ 10 minute, preferably 2 ~ 7 minutes, as 5 minutes.

The present inventor studies discovery, and the fragrance matter in passion fruit can at high temperature volatilize lost, therefore preferred in the method for first aspect present invention, and the intensification of step (3) and/or the cooling of step (4) are carried out in closed container.

The present inventor studies discovery in addition, if fermentate is too much or very few relative to closed container, capital affects final passion fruit juice quality, so preferred in the method for first aspect present invention further, the fermentate of step (3) and/or the volume of fermentate of step (4) and the volumetric ratio of closed container are 1: 1.05 ~ 1.8, be preferably 1: 1.1 ~ 1.5, be more preferably 1: 1.2 ~ 1.3, most preferably be 1: 1.25.

In addition, preferably in the method for first aspect present invention, in step (4), cooling can be Temperature fall (cooling), also can be to accelerate (as with low-temperature receivers such as refrigerators, or water-cooled) cooling.

The present inventor studies discovery, the main advantage of passion fruit fruit juice (comprising the pure passion fruit fruit juice of prior art) is fragrance, taste meta-acid, do not attract all groups, especially children, so in the method for preferred first aspect present invention, in step (5), the filtrate obtained to step (4) adds water, sugared and/or Other Drinks, especially sugaring and/or Other Drinks.And, the present inventor studies discovery, and sugaring etc. inhibit tart flavour, is not suitable for the crowd of having good taste pursuing mouthfeel stereovision, but but can attract the crowds such as children, therefore can determine according to the target group of corresponding beverage that optional project selects or do not select.

In second aspect, fruit juice prepared by the method that the invention provides first aspect present invention.This fruit juice can be preserved for a long time and never degenerate, and except original passion fruit local flavor, fragrance and taste can increase stereovision further, and be particularly suitable for the crowd that has good taste pursuing stereovision, the product orientation usually for this part crowd is higher, and added value is also higher.

In the third aspect, the invention provides esterase and preparing the application in passion fruit fruit juice.This to prepare in passion fruit fruit juice first Application esterase.

Preferably in the application of third aspect present invention, esterase is the preferred esterase of first aspect present invention, i.e. the amino acid sequence that has of esterase:

(1) as shown in SEQ ID NO:2;

(2) amino acid sequence of reservation esterase active amino acid sequence SEQ ID NO:2 being increased, lack and/or replace one or several amino acid residue and obtain.

The present invention has following advantages and effect: can carry out globality process to the whole fruit of passion fruit, carries out getting juice process after can isolating the pulp of band seed, even separation seed as prior art again; Sharp sword can be used to pulverize, and equipment is conventional and cost is low, and operational means is easy, is also convenient to automation; Passiflora edulis fruit juice beverage and cake shape food can be obtained simultaneously; Passion fruit fruit juice is not easy by the oil interference quality in seed, long shelf-life; No matter passion fruit fruit juice is fragrance or taste, and have levels sense all more, is particularly suitable for towards high-end consumer groups, increases value-added content of product.

For the ease of understanding, below will be described in detail the present invention by specific embodiment.It is important to note that instantiation is only to illustrate, do not form limitation of the scope of the invention.Obvious those of ordinary skill in the art according to illustrating, can make various correction and change to the present invention herein within the scope of the invention, and these are revised and change and also include in scope of the present invention.In addition, the present invention refer to open source literature, and these documents are also to more clearly describe the present invention, and their entire contents is all included the present invention in and carried out reference, just looks like that repeated description is excessively the same in the description of the present invention for their full text.

Summary of the invention

The technical problem to be solved in the present invention is to provide the preparation technology of new passion fruit fruit juice.In addition, present invention also offers the esterase etc. with using in the passion fruit fruit juice of this explained hereafter and this technique.

Specifically, in first aspect, the invention provides the preparation method of passion fruit fruit juice, it comprises:

(1) passion fruit cleaned is pulverized;

(2) crushed material obtained to step (1) adds water and esterase fermentation;

(3) fermentate that step (2) obtains is heated up;

(4) to fermentate cooling (being preferably reduced to room temperature) that step (3) obtains, then filter; With,

(5) filtrate optionally obtained to step (4) adds water, sugared and/or Other Drinks.

In the present invention, pulverizing can make the seed of passion fruit broken, and grease wherein can be mixed into final passion fruit fruit juice, but the present inventor innovatively overcomes such defect by adding the means such as esterase.Preferably in the method for first aspect present invention, in step (1), be crushed to the largest particles diameter and be no more than 0.5cm.Generally for the cost saved and pulverize, there is no need to pulverize thorough especially, can stop when the largest particles diameter is no more than 0.5cm pulverizing, so the largest particles of diameter more than 0.3cm, even 0.4cm can be there is in crushed products.

detailed description of the invention

Below by specific embodiment, invention will be described herein.As do not specialized part, can according to those skilled in the art in the brewing technology be familiar with and gene recombination technology handbook and the bibliography quoted herein listed method implement.In addition, the material and facility used in embodiment all can be commercially.

The preparation of embodiment 1 esterase

According to the gene order of our Commission Design (see SEQ ID NO:1, the amino acid sequence of esterase of its coding is SEQ ID NO:2), entrust Guangdong Jida Genetic Pharmaceutical Engineering Research Center Co., Ltd. to synthesize this esterase gene by commercial sources and be cloned into yeast cells and be built into secreting, expressing bacterial strain.Cloning procedure can carry out with reference to " Molecular Cloning: A Laboratory guide ", in brief, synthesize the fragment of this gene, splicing, take splicing product as template, pcr amplification is carried out in 50 μ L reaction volumes, wherein forward primer is as shown in SEQ ID NO:3, reverse primer is as shown in SEQ ID NO:4, collect the amplified production of about 1.4kb, with EcoR I and .Xba I double digestion, then be connected on the pPICZ α A (Invitrogen) of same double digestion, positive construct electricity is transformed in yeast GS115 strain (Invitrogen), positive yeast is filtered out with Zeocin, after through pcr amplification, also order-checking confirms to contain the correct gene order as shown in SEQ ID NO:1, return to us.

Positive yeast is inoculated in 25ml seed culture medium, cultivate in 30 DEG C with 220rpm concussion, until OD600 reaches 3.5, wherein the formula of seed culture medium is: 2% (W/W) peptone, 1% (W/W) yeast extract, 1.34% (W/W) YNB (without amino yeast nitrogen), 4*10 ^-5% (W/W) biotin and pH3.2 sodium hydrogen phosphate-citrate buffer solution (0.2M sodium hydrogen phosphate: 0.1M citric acid (volume ratio)=1: 3).Then, 25ml seed culture product is inoculated in 100ml fermentation medium, cultivate in 30 DEG C with 200rpm concussion, within every 12 hours, add methyl alcohol to 0.5% (V/V), cultivate 96 hours, wherein the formula of fermentation medium is: 2% (W/W) peptone, 1% (W/W) yeast extract, 1.34% (V/V) YNB (without amino yeast nitrogen), 4*10 ^-5% (W/W) biotin, 0.5% (V/V) methyl alcohol and pH3.2 sodium hydrogen phosphate-citrate buffer solution.Then, in 4 DEG C with the centrifugal fermented and cultured product of 1500rpm, retain supernatant and be splined on Sephadex G-75 chromatographic column, wash-out is carried out with pH3.2 sodium hydrogen phosphate-citrate buffer solution, collect the maximum elution peak that 280nm UV detect goes out, then freeze drying, in 4 DEG C of preservations, namely prepares esterase of the present invention.After testing, esterase expression is 3.7g/L.

The preparation of embodiment 2 fruit juice

Select maturation, without putrefaction, qualified passion fruit without mildew, clean, directly whole fruit cutter is shredded, shred and stop when the largest particles diameter is no more than 0.5cm.All chopping products (containing fluid and solid) are loaded in the fermentation tank with agitator arm in the lump, add the distilled water of this products weight 1.2 times, and add the esterase prepared according to the method for embodiment 1 to 0.05% of product and water gross weight, after stirring, stir fermentation 24 hours in 35 DEG C with 50rpm.Then, tunning is loaded with in the sterilization tank of agitator arm, fill after 80% of sterilization tank internal volume, covered and enclosed sterilization tank, heat and stir thus tunning entirety is evenly heated up with the speed of about 10 DEG C/min, during to 85 DEG C, reducing thermal source with this temperature 5 minutes, then remove thermal source, naturally cool to room temperature.Then open the lid of sterilization tank, aseptically filter the sterilizing product of acquisition, retain filtrate (filter cake sweet and sour taste, seed is also softening, filter residue (after optional sugaring) can be prepared becomes cake shape food), namely passion fruit fruit juice, canned.

In addition, in contrast, replace esterase of the present invention with the high-quality pectase of Novozymes Company (PECTINEX Ultra SP-L), carry out above step, obtain reference substance.

The inspection of embodiment 3 fruit juice

The passion fruit fruit juice prepared according to the method for embodiment 2 and reference substance thereof to be placed in 42 DEG C of environment 7 days, carry out accelerated test, find, visually, passion fruit fruit juice is homogeneous, clarification, without layering, and reference substance has lamination, aqueous phase has one deck grease, and the muddy naked eyes of aqueous phase can be distinguished; In sense of smell, passion fruit fruit juice aromas is pleasant, by professional product, fragrant teacher has told outside the banana that should have, pineapple, strawberry, lichee, lemon and mango fragrance, also find that there is slight peanut fragrance, reference substance then has dense peculiar smell, therefore stops and taste test fragrant to the news of reference substance; In the sense of taste, passion fruit fruit juice, except original sour-sweet taste, also has slight tannin taste, wherein tart flavour and tannin taste suppressed after sugaring.

Claims (10)

1. the preparation method of passion fruit fruit juice, it comprises:

(1) passion fruit cleaned is pulverized (preferred powder is broken to the largest particles diameter and is no more than 0.5cm);

(2) crushed material obtained to step (1) adds water and esterase fermentation;

(3) fermentate that step (2) obtains is heated up;

(4) to fermentate cooling (being preferably reduced to room temperature) that step (3) obtains, then filter; With,

(5) filtrate optionally obtained to step (4) adds water, sugared and/or Other Drinks.

2. method according to claim 1, wherein in step (1), pulverizing is pulverized with sharp sword (e.g., cutter).

3. method according to claim 1, wherein, the weight ratio of the passion fruit in the water in step (2) and step (1) is 0.8 ~ 2: 0.5 ~ 1.5, is preferably 1 ~ 1.5: 0.8 ~ 1.2, is more preferably 1.1 ~ 1.3: 0.9 ~ 1.1.Most preferably be 1.2: 1.

4. method according to claim 1, wherein in step (2), the amino acid sequence that esterase has:

(1) as shown in SEQ ID NO:2;

(2) amino acid sequence of reservation esterase active amino acid sequence SEQ ID NO:2 being increased, lack and/or replace one or several amino acid residue and obtain.

5. method according to claim 1, wherein in step (3), the temperature reached that heats up is 80 ~ 95 DEG C, preferably 82 ~ 90 DEG C, as 85 DEG C; And/or, the temperature that keeping heats up reaches 1 ~ 10 minute, preferably 2 ~ 7 minutes, as 5 minutes.

6. method according to claim 1, wherein, the intensification of step (3) and/or the cooling of step (4) are carried out in closed container.

7. method according to claim 6, wherein, the fermentate of step (3) and/or the volume of fermentate of step (4) and the ratio of the volume of closed container are 1: 1.05 ~ 1.8, are preferably 1: 1.1 ~ 1.5, be more preferably 1: 1.2 ~ 1.3, most preferably be 1: 1.25.

8. the fruit juice prepared of the arbitrary described method of claim 1 ~ 7.

9. esterase is preparing the application in passion fruit fruit juice.

10. application according to claim 9, the wherein amino acid sequence that has of esterase:

(1) as shown in SEQ ID NO:2;

(2) amino acid sequence of reservation esterase active amino acid sequence SEQ ID NO:2 being increased, lack and/or replace one or several amino acid residue and obtain.