Coffee Fermentation Research Articles

Technology innovation for production of specialty coffee

Coffee is one of the most important agricultural commodities in the world. Data from International Coffee Organization (ICO) indicate that total world production increased from 8.945 million metric tons in 2014 to 9.126 million metric tons in 2015, 9.462 million metric tons in 2016, and 9.580 million metric tons in 2017. Data from ICO also show that coffee is commercially cultivated in 56 countries and Indonesia with total production of about 720 thousand tons in 2017 was the fourth largest producer after Brazil (3.06 million tons), Vietnam (1.77 million tons), and Colombia (840 thousand tons). To increase the economic value of this commodity, the quality of coffee bean must be improved through both agronomic and postharvest aspects. This study was designed to develop a new ohmic-based fermentation technology for coffee cherry to improve flavor quality of coffee beans. Results of this study indicate that this technology can produce coffee with cup-test scores ranging from 84.38 to 86.88 with an average score of 85.713. This score is higher than the cup test score of Indonesia civet coffee (luwak coffee) reported by several researchers. Therefore, application of ohmic heating based technology for coffee fermentation can significantly improve flavor quality of coffee beans.

Kualitas Organoleptik dan Nilai pH Kulit Kopi yang Difermentasi dengan Penambahan Level Trichoderma sp. yang Berbeda

This study aimed to determine the quality of organoleptic and pH value of fermented coffee peel using Trichoderma sp. at different levels. The treatment of coffee peel fermented using Trichoderma sp. consisted of P0 (without Trichoderma sp.), P1 (7.5 g), P2 (15.0 g), and P3 (22.5 g). Each treatment consisted of 4 replications. The parameters observed included: organoleptic quality (color, aroma, and texture) and pH value. The organoleptic test using the scoring method by 12 panelists. The result was analyzed by analysis of variance (ANOVA). A comparison among different treatments was performed by Duncan Multiple Range Test (DMRT). Fermentation of coffee peel waste with Trichoderma sp. produces average light brown color, sour aroma, and softer texture with an increasing percentage of Trichoderma sp. The pH value of coffee peel fermentation with Trichoderma sp. ranged around 6.

Kualitas Organoleptik dan Nilai pH Kulit Kopi yang Difermentasi dengan Penambahan Level Trichoderma sp. yang Berbeda

This study aimed to determine the quality of organoleptic and pH value of fermented coffee peel using Trichoderma sp. at different levels. The treatment of coffee peel fermented using Trichoderma sp. consisted of P0 (without Trichoderma sp.), P1 (7.5 g), P2 (15.0 g), and P3 (22.5 g). Each treatment consisted of 4 replications. The parameters observed included: organoleptic quality (color, aroma, and texture) and pH value. The organoleptic test using the scoring method by 12 panelists. The result was analyzed by analysis of variance (ANOVA). A comparison among different treatments was performed by Duncan Multiple Range Test (DMRT). Fermentation of coffee peel waste with Trichoderma sp. produces average light brown color, sour aroma, and softer texture with an increasing percentage of Trichoderma sp. The pH value of coffee peel fermentation with Trichoderma sp. ranged around 6.

The Fermentation of Parchment Ohmic Technology Used to Process Coffee Beans (A Wet Black Coffee) Aroma

This study aimed (1) to determine the effect of temperature on and (2) to determine the fermentation time and the effect of bromelin enzyme of coffee beans aroma on). The process of fermentation of coffee beans using the ohmic technology with the addition of pineapple extract. The treatment used in the fermentation with Ohmic technology was the temperatures of 35°C dan 45°C; and fermentation times were 6, 12, and 18 hours. The observation parameters included pH, caffeine content, and organoleptics. The research results indicated that in the fermentation of the coffee beans using Ohmic technology at the temperature of 35°C and the time fermentation of 12 hours, the caffeine content (1.6%) became 0.047%, and the temperature of 35°C and the time of fermentation was 12 hours had produced pH of 4.81, and the temperature of 45°C with the time fermentation of 6 hours, high caffeine value of 0.147%, and at 45°C with the time fermentation of 18 hours had produces pH value of 5.83. The organoleptic description of the flavor attributed with temperature of 35°C, the temperature flavor attributed and the time fermentation of 12 hours, the average body score was 4.94, while the bitterness was 3.7, and at the temperature of 45°C and the time fermentation of 12 hours produced the body weight scores of 3.49, while the bitterness of 3.45. The fermented coffee beans with the clinical technology was very effective to use in.ohmci technology because it could provide a relatively uniform and evenly distributed heating system.

Isolation and characterization of yeast isolated from civet (Paradoxorus hermaphroditus)

Civet coffee is one of the most popular coffee in the world. This coffee is fermented naturally in the digestive tract of civet (Paradoxorus hermaphroditus). The coffee fermentation is characterized by the presence of different microorganisms belonging to the groups of yeast, fungi and bacteria. The aim of this work were to isolation and characterization of yeast isolated from civet. The isolation was carried out using Sabouraud Dextrose Agar (SDA). A total of three different yeasts (YC1, YC2 and YC3) were isolated and subjected to morphology and biochemical test. The result showed that all isolates had various characteristics on carbohydrate fermentation, temperature tolerance and proteolityc activity. Two isolates (YC1 and YC2) showed good proteolityc activity. Further investigation is needed to identify and select of yeast performance on coffee fermentation.

Isolation, Identification, and Characterization of Pectinolytic Yeasts for Starter Culture in Coffee Fermentation.

This experiment was carried out to identify and select pectinolytic yeasts that have potential use as a starter culture for coffee fermentation during wet processing. The coffee fruit was fermented for 48 h at 28 °C and a sample was taken from the fermented solution and spread onto yeast extract-peptone-dextrose agar (YPDA) media and incubated at 28 °C. A total of 28 yeasts were isolated, eight of which had the ability to produce pectinase enzymes. The species of those eight yeasts were molecularly identified and confirmed. These yeasts are Wickerhamomyces anomalus (strain KNU18Y3), Saccharomycopsis fibuligera (strain KNU18Y4), Papiliotrema flavescens (strain KNU18Y5 and KNU18Y6), Pichia kudriavzevii (strain KNU18Y7 and KNU18Y8), and Saccharomyces cerevisiae (strain KNU18Y12 and KNU18Y13). The pectin degradation index of S. fibuligera (strain KNU18Y4), W. anomalus (strain KNU18Y3), and P. flavescens (strain KNU18Y6) were higher compared to the others, at 178%, 160%, and 152%, respectively. The pectinase enzyme assays were made on two growth media: coffee pulp media (CPM) and synthetic pectin media (SPM). S. fibuligera (strain KNU18Y4) and W. anomalus (strain KNU18Y3) had great potential in producing polygalacturonase (PG) and pectin lyase (PL) compared to others in both media. However, S. cerevisiae strains (KNU18Y12 and KNU18Y13) produced higher pectin methylesterase (PME). Using MEGA 6 software, the phylogenetic trees were constructed to determine the evolutionary relationship of newly identified yeasts from our experiment and previously published yeast species. The sequences of the yeasts were deposited in the National Center for Biotechnology Information (NCBI) database.

Effect of Co-Inoculation with Pichia fermentans and Pediococcus acidilactici on Metabolite Produced During Fermentation and Volatile Composition of Coffee Beans

Removal of the mucilage layer of coffee fruits by a fermentation process has became an interesting strategy to improve coffee quality, which is able to assist the formation of flavored molecules. In this study, four sets of inoculation protocols were evaluated using ripe and immature coffee fruits, respectively, including (i) pure culture fermentation with Pichia fermentans, (ii) pure culture fermentation with Pediococcus acidilactici, (ii) combined fermentation with P. fermentans and P. acidilactici, and (iv) spontaneous, non-inoculated control. The initial pulp sugar concentration of ripe coffee fruits (0.57 and 1.13 g/L glucose and fructose content, respectively) was significantly higher than immature coffee pulp (0.13 and 0.26 g/L glucose and fructose content, respectively). Combined inoculation with P. fermentans and P. acidilactici of ripe coffee beans increased pulp sugar consumption and production of metabolites (lactic acid, ethanol, and ethyl acetate), evidencing a positive synergic interaction between these two microbial groups. On the other hand, when immature coffee fruits were used, only pure culture inoculation with P. fermentans was able to improve metabolite formation during fermentation, while combined treatment showed no significant effect. Altogether, 30 volatile compounds were identified and semi-quantified with HS- solid phase microextraction (SPME)-gas chromatography coupled to mass spectrophotometry (GC/MS) in fermented coffee beans. In comparison with pure cultures and spontaneous process, combined treatment prominently enhanced the aroma complexity of ripe coffee beans, with a sharp increase in benzeneacetaldehyde, 2-heptanol, and benzylalcohol. Consistent with the monitoring of the fermentation process, only P. fermentans treatment was able to impact the volatile composition of immature coffee beans. The major impacted compounds were 2-hexanol, nonanal, and D-limonene. In summary, this study demonstrated the great potential of the combined use of yeast and lactic acid bacteria to improve fermentation efficiency and to positively influence the chemical composition of coffee beans. Further studies are still required to investigate the mechanisms of synergism between these two microbial groups during the fermentation process and influence the sensory properties of coffee products.

Coffee starter microbiome and in-silico approach to improve Arabica coffee

Arabica coffee is fermented to obtain intense flavor profile and finds elusive prospects in the coffee industry. The Arabica coffee mucilage constitute 2–5% of the fruit dry weight with 94% moisture, 4% sugar, 0.7% protein and 1–3% pectin. Inductive isolation and screening of functional attributes of microbial strains were examined. Fermentative vigor along with the enzymatic progression of the microbes was streamlined and the potential isolates of consortia were optimized. The In-silico docking studies on the fermentation mechanism evidenced minimal interaction between Pectin and pectinase at lower energy level. The 1CZF disclose the best interaction binding energy (−3.92 kJ mol−1) with rapid enzymatic and desired fermentation. A CCRD was employed to study the interactive effect of isolates. The consortia of Saccharomyces cerevisiae, Lactobacillus plantarum and Bacillus sphaericus (1:1:1) at 10% inoculum concentration found significant in demucilizatin of coffee beans with noticeable improvement in alcohol (70.26 mg/ml), sugar (5.5 mg/ml) and pectinase (11.66 U/ml) compared to natural fermentation. The sensory profile of starter fermention(G) scored 7.0 on 1–10 scale. Prospective insights on application of starter consortia on Arabica coffee fermentation indicates prime requisite for coffee industry.

Coffee mucilage impact on young coffee seedlings and soil microorganisms

AbstractA greenhouse pot experiment was carried out to assess the effects of fermented coffee mucilage applied as mulch together with maize leaves on the growth of young coffee plants of two different varieties and on soil microbial biomass indices. The coffee variety Catuai required 32% more water per g plant biomass than the variety Yellow Caturra, but had a 49% lower leaf area, 34% less shoot and 46% less root biomass. Maize and mucilage amendments did not affect leaf area, shoot and root yield, or the N concentration in shoot and root dry matter. The amendments always reduced the water use efficiency values, but this reduction was only significant in the maize+mucilage‐14 (= 14 g mucilage pot−1) treatment. Soil pH significantly increased from 4.30 in the control to 4.63 in the maize+mucilage‐14 treatment. Microbial biomass C increased by 18.5 µg g−1 soil, microbial biomass N by 3.1 µg g−1 soil, and ergosterol by 0.21 µg g−1 soil per g mucilage added pot−1. The presence of mucilage significantly reduced the microbial biomass‐C/N ratio from a mean of 13.4 in the control and maize treatments to 9.3, without addition rate and coffee variety effects. The application of non‐composted mucilage is recommended in areas where drought leads to economic losses and in coffee plantations on low fertility soils like Oxisols, where Al toxicity is a major constraint.

First description of bacterial and fungal communities in Colombian coffee beans fermentation analysed using Illumina-based amplicon sequencing

In Colombia, coffee growers use a traditional method of fermentation to remove the cherry pulp surrounding the beans. This process has a great influence on sensory quality and prestige of Colombian coffee in international markets, but has never been studied. Here we use an Illumina-based amplicon sequencing to investigate bacterial and fungal communities associated with spontaneous coffee-bean fermentation in Colombia. Microbial-derived metabolites were further analysed by high–performance liquid chromatography and gas chromatography–mass spectrometry. Highly diverse bacterial groups, comprising 160 genera belonging to 10 phyla, were found. Lactic acid bacteria (LAB), mainly represented by the genera Leuconostoc and Lactobacillus, showed relative prevalence over 60% at all sampling times. The structure of the fungal community was more homogeneous, with Pichia nakasei dominating throughout the fermentation process. Lactic acid and acetaldehyde were the major end-metabolites produced by LAB and Pichia, respectively. In addition, 20 volatile compounds were produced, comprising alcohols, organic acids, aldehydes, esters, terpenes, phenols, and hydrocarbons. Interestingly, 56 microbial genera, associated with native soil, seawater, plants, insects, and human contact, were detected for the first time in coffee fermentation. These microbial groups harbour a remarkable phenotypic diversity and may impart flavours that yield clues to the terroir of Colombian coffees.

Antioxidant Activity, Total Polyphenol, Flavonoid and Tannin Contents of Fermented Green Coffee Beans with Selected Yeasts

We examined the antioxidant activity, total polyphenol content (TPC), total flavonoid content (TFC), total tannin content (TTC) and physical characteristics of green coffee beans fermented with selected yeasts. There was no significant (p > 0.05) interaction effect between yeast-fermented coffee extracts and duration of fermentation on antioxidant activity (oxygen radical absorbance capacity [ORAC] and superoxide dismutase-like [SOD-like] activity). However, the mean of the antioxidant activity (ORAC and SOD-like activity) significantly (p < 0.05) increased in the fermented coffee extracts compared to unfermented coffee. There were significant (p < 0.05) interaction effects between yeast-fermented coffee extracts and duration of fermentation (24 h and 48 h) on the TPC, TFC, TTC and pH of the fermented solution and on the colors of the ground-roasted coffee. The TPC showed a pattern of increase in samples Ferm-1 and Ferm-3 as fermentation time increased from 24 h to 48 h. However, a decreasing TPC trend was observed in Ferm-2 as the number of fermentation hours increased from 24 to 48. The fermented coffee beans had a significantly higher flavonoid content than the unfermented coffee beans, while fermentation significantly decreased the tannin content compared to that in unfermented coffee.

The Role of Microbes in Coffee Fermentation and Their Impact on Coffee Quality

Coffee is one of the most important and widely used commercial crops in the world. After ripe coffee cherries are harvested, coffee must pass through several steps to become (green) raw coffee beans. Commonly, there are three different processing methods used to obtain green coffee beans from coffee cherries, namely, the wet, dry, and semidry methods. Microorganisms (yeasts and bacteria) play a major role in coffee fermentation process by degrading mucilage by producing different enzymes (pectinase), acids, and alcohols. Starter culture development is crucial and is done by selecting microorganisms that have certain characteristics, such as mucilage degradation ability, tolerance to stress during fermentation, the ability to suppress the growth of pathogenic fungi, and a positive impact on the sensory quality of the coffee. Currently, green coffee beans obtained from farms that use any of the above processing methods are fermented with selected microorganisms to improve the flavour and aroma of the coffee. This is the result of a new insight into the development of unique flavoured coffee and into engaging with the coffee market to better benefit. This review gives a comprehensive overview of the fermentation process, microorganisms and starter cultures, and fermentation’s impact on coffee quality. Future prospects are also discussed through the incorporation of recent research.

Bioactivity of Fermented Green Coffee Bean Extract Containing High Chlorogenic Acid and Surfactin

Chlorogenic acid (CGA) is a major component of green coffee beans. Surfactin, a cyclic lipopeptide, is produced and secreted by Bacillus subtilis strains. In this study, bioactivities of fermented green coffee bean extract (FGCBE) and the individual compounds, CGA and surfactin. were compared in HepG2 cells. The concentration of surfactin and CGA in the FGCBE and non-fermented green coffee bean extract (NFGCBE) were determined to be 9.2 and 7.33 and 0.72 and 0.53 mg·mL-1, respectively. The FGCBE contained about 20% and 26% more CGA and surfactin than the NFGCBE. Although CGA and surfactin exhibited cytotoxicity at concentrations more than 100 and 20 μg respectively, the FGCBE 50 containing CGA (460 μg·mL-1) and surfactin (720 μg·mL-1) effectively prevented cell death by oxidative stress and also strongly activated the proliferation of cells incubated with under 50 μM H2O2. The CGA and surfactin in FGCBE were 9.2 and 72 times higher than the CGA and surfactin compounds (50 and 10 μg·mL-1). The relative proliferation of the FGCBE-treated cells also was 3.3 and 8.8 times higher than the CGA and surfactin compounds treated the oxidative stressed cells with 50 μM H2O2. These results suggest that the single compounds such as CGA and surfactin generally have cytotoxicity at low concentration of them but FGCBE contained them acted as strong antioxidants, activators of cell proliferation, inhibitors of cell apoptosis. Various bioactive compounds in fermented coffee bean also seem to help cell proliferation and decreasing of cytotoxicity by CGA and surfactin in coffee bean.

Luwak coffee in vitro fermentation : literature review

Luwak coffee is one of the Indonesia traditional coffee that known globally because of its unique aroma and flavor. Luwak coffee was produced by a fermentation process in luwak digestion by involving microbe that can be found in luwak digestion. Luwak coffee fermentation process using luwak animal has many weakness such as inefficient production processes, and final product quality that always makes consumers hesitate in terms of its hygiene and halal. Therefore some research to produce coffee with the quality of luwak coffee without using luwak began to be developed. Several methods of fermentation to produce coffee with quality luwak coffee are discussed in this paper, such as fermentation by in vitro technique

Sensory Profile of Fermented Arabica Coffee in the Perception of American Cupping Tasters

The consumption of specialty coffees has been growing year after year, indicating that several niches and market forms according to the sensorial orientation of the consumers, as well as the cupping tasters and agents are responsible for the composition of blends, and for the physical and sensory classification of the coffees in the industry. This study presented 5 methods of spontaneous and induced fermentation to different North American cupping tasters, in order to evaluate coffee quality potential, based on the Specialty Coffee Association (SCA) protocol. During the months of May to July of 2017, 150 coffee samples were evaluated. Initial results indicate that North American cupping tasters evaluated dry fermented coffees using yeast culture as superiors, in the sensorial aspect, than the more conventional methods in Brazil. Such indication opens new discussions on the acceptance of a determined product, followed by the potential for modification of the sensory route as of the fermentation that will affect the attributes.

Formulation and Storage Stability of Nanoencapsulated Proteus penneri and Bacillus aerophilus

Proteus penneri and Bacillus aerophilus are bacterial that can be used as dry starter in the fermentation of artificial civet coffee. The use of dried starter will facilitate the handling, distribution and transportation of culture. The aims of this research were to evaluate the suitable nanoencapsulation coating materials of Proteus penneri and Bacillus aerophilus to increase biomass cells during drying and storage. Nanoencapsulated starter using carrier materials maltodextrin, sodium alginate and skim milk with spray drying method. The Nanoencapsulated Proteus penneri and Bacillus aerophilus were tested for stability during storage for up to 14 days at different temperatures (4 and 37 °C). Proteus penneri isolates had higher viability when encapsulated using maltodextrin with a concentration of 30%. Bacillus aerophilus isolates had higher viability when encapsulated using maltodextrin at 15% concentration. Viability of Bacillus aerophilus and Proteus penneri in dry starter with filler coffee pulp powder and coffee bean powder was less than after nanoencapsulation. The storage stability studies demonstrated the storage time and temperature affecting the survival of cells of dried starter cultures.

Microbiological and chemical-sensory characteristics of three coffee varieties processed by wet fermentation

This work evaluated the bacterial diversity during coffee wet fermentation of the three coffee varieties—Mundo Novo (MN), Ouro Amarelo (OA), and Catuai Vermelho (CV). Isolates were identified by polyphasic techniques: biochemical tests, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and DNA sequencing. Chemical compositions were determined by high (HPLC) and gas chromatography-mass spectrometry (GC-MS) and the roasted beans were sensorial evaluated using the cupping test. Thirty-six mesophilic bacteria and six lactic acid bacteria were identified. Lactobacillus plantarum and Leuconostoc mesenteroides were often found in all varieties. Citric acid was the acid detected in higher concentrations. The volatile profile of the green coffee beans changed during the fermentation in the tank, but more significantly, during the roasting process. These volatiles belonged to the classes of acids, alcohols, aldehydes, and hydrocarbons. Temporal dominance of sensations analysis showed sensorial sensations of acidity (OA and CV), bitterness, chocolate, nuts (MN), and sweetness (CV). The characteristics of each coffee variety were distinct, mainly in relation to total bacteria population, volatile compounds, and sensorial profile. In conclusion polyphasic methodology was efficiently done for bacteria identification; the dominant bacteria might be used for starter cultures and the chemical and sensory analyses helped to understand the changes in coffee fermentation. Our findings are relevant to future select starter bacteria for coffee processing to improve quality and standardization of quality.

“CIRAGI”- The microbial fermentation starter for developing excellent coffee flavour

Most of the green bean coffees produced by smallholders are not properly fermented, while green bean coffees from large plantations undergo a spontaneous fermentation involving present microbial activity in the processing environment. The process of coffee fermentation itself involves not less than 144 endogenous microbial species originating from coffee fruits or from the surrounding environment. This condition is the source of inconsistency and low cup quality of coffee beans. The inoculation of selected microbial fermentation starter can transform microbial ecosystem. The transformation leads to beneficial microbes dominating and actively inducing flavour precursors into excellent aroma during roasting. The Indonesian Research Institute for Biotechnology and Bioindustry (IRIBB) has developed the CIRAGI (abbreviation of CItaRAsa tingGI meaning “a high flavour” in Indonesian), a fermentation starter for coffee beans using selected Lactobacillus sp. as the active ingredient. High cup quality coffee can consistently result from well-controlled fermentation process of the coffee bean using CIRAGI. The fermentation of both Robusta and Arabica coffee beans using CIRAGI has been demonstrated at laboratory and field trials. The cup tests of both Arabica and Robusta coffee have resulted in improved quality of the coffee flavour, increase of cup test scores based on the SCAA parameter, and elimination of off-flavour.

Enhancement of antioxidant properties from green coffee as promising ingredient for food and cosmetic industries

Abstract Phenolic compounds are present in large quantities in green coffee and have been recognized as natural antioxidants. However, hydroxycinnamic acids such as chlorogenic acids are found mainly in the esterified form which reduces their bioavailability. Therefore, the aim of this study was to perform the biotransformation of the green coffee using Aspergillus oryzae. Green coffee was fermented under solid state at 25 °C for 24, 48 and 72 h and the phenolic compounds were extracted using a hydroethanolic solution. The antioxidant activity of the extracts was evaluated by the 2,2-diphenyl-1-picrilhidrazyl and reducing power methods, and the determination of total phenolic contents was performed using Folin-ciocalteu method. Chlorogenic acid (5-ACQ), caffeic acid and caffeine were quantified by HPLC. Green coffee extracts fermented for 24 h presented an increase of 115.7% and 66.4% of the antioxidant activity measured by DPPH• and reducing power methods, respectively. Additionally, coffee fermentation promoted an increase of 68.6% in the content of phenolics, and an increase of chlorogenic and caffeic acids. The results showed the biotransformation as a strategy to obtain an enriched extract of antioxidant compounds as promising ingredient for food and cosmetic industries.

Sensory and Microbial Characteristics of Civet Coffee

Civet coffee has been known as the most expensive coffee in the world. Its high prices are mainly contributed by its uncommon production method. Civet coffee is produced from the coffee beans which have been digested by Asian palm civet or Luwak ( Paradoxorus hermaprodites ). Civet coffee production is very limited due to its processing is merely depends on civet biologic system. Unfortunately, reliable information on civet microbial digestive is still limited. The aim of this research is to evaluate cup (sensory) and microbial quality of wild and caged civet coffee. Samples of wild and caged civet coffee obtained from Bener Meriah District, Aceh, Indonesia. Sensory quality parameters consist of fragrance, aroma, flavour, aftertaste, acidity, body, balance, uniformity, clean cups, sweetness and overall. Microbial quality was done by enumeration of lactic acid bacteria. The results showed that wild and caged civet coffee relatively had similar profile sensory and microbial characteristics. The score cupping of wild civet coffee is slightly higher than caged civet coffee. Aroma and flavour characteristics of wild and caged civet coffee were herby, nutty, grassy tobacco-like, fishy and sweet but there were over fruity-papaya and guava-like aroma and flavour of caged civet coffee. The lactic acid bacteria count on wild and caged civet were 5.8 x 10 8 cfu/ml and 3,8 x10 8 cfu/ml. Three different colonies from MRS media were isolated and submitted to gram and catalase test. All isolates showed gram-positive and catalase negative. The result confirmed that most colonies are lactic acid bacteria group. Two isolates (ICF 1 and ICF 3) showed proteolytic activity, and isolated ICF 1 showed the largest zone of clearance of 15 mm. Further investigation is required for identification and selection of performance lactic acid bacteria for artificial civet coffee fermentation.